摘要
为建立农杆菌介导的朝仓花椒遗传转化体系,以朝仓花椒叶柄及茎段为转化受体,研究了预培养时间、侵染菌液浓度、侵染时间、共培养时间等因素对转化效率的影响。获得转化体系如下:外植体预培养3d,在OD600为0.5～0.8菌液侵染10min,共培养3d,延迟7d筛选,以30和50mg/L卡那霉素(Kan)进行梯度筛选。结果表明:重悬液及共培养基中添加100μmol/L乙酰丁香酮(AS),可提高朝仓花椒Kan抗性愈伤率至24.6%;转化植株经GUS组织活性及PCR检测呈阳性,证实外源基因已导入朝仓花椒基因组中,利用本转化体系可为下一步将有重要经济价值外源基因导入花椒打下基础。
In order to establish an Agrobacterium-mediated genetic transformation system for Zanthoxylum piperitum DC.var.inerme Makino,using petioles and stems as explants,we examined several influencing factors of transformation efficiency,including pre-cultivation time,optimum bacterium concentration and inflection period,and co-cultivation time.The efficient transformation system obtained was 3 days pre-cultivation time,10 minutes inflection using bacterium at the concentration of OD600 0.5-0.8,followed by 3 days co-cultivation.Selection of the transgenic plants was carried out after 7 days regeneration using gradient kanamycin at 30 and 50 mg/L,respectively.Adding 100 μmol/L acetosyringone in the re-suspending solution and co-cultivation medium increased the transformation efficiency to 24.6%.Transgenic plants were confirmed by GUS histochemical assays and polymerase chain reaction.The results suggest that an efficient Agrobacterium-mediated transformation system for stable integration of foreign genes into Z.piperitum DC.var.inerme Makino has been developed,which could be applied in future studies on transferring economically important genes into Z.pipertum DC.var.inerme Makino.
出处
《北京林业大学学报》
CAS
CSCD
北大核心
2011年第6期80-85,共6页
Journal of Beijing Forestry University
基金
国家转基因新品种培育重大专项(2008ZX08010-003)
关键词
朝仓花椒
农杆菌
遗传转化
Zanthoxylum piperitum DC.var.inerme Makino
Agrobacterium tumefaciens
genetic transformation
