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农杆菌介导法将LFY基因导入苹果的研究
引用本文:刘静,邵建柱,徐继忠,李湘利,郭文武.农杆菌介导法将LFY基因导入苹果的研究[J].河北农业大学学报,2006,29(3):6-9.
作者姓名:刘静  邵建柱  徐继忠  李湘利  郭文武
作者单位:1. 河北农业大学,园艺学院,河北,保定,071001
2. 济宁师范专科学校,生物系,山东,济宁,272025
3. 华中农业大学,作物遗传改良国家重点实验室,湖北,武汉,430070
基金项目:河北省科技厅科技攻关项目 , 河北农业大学校科研和教改项目
摘    要:以M26苹果叶片为外植体,利用GUS瞬时表达率,对农杆菌介导法转化苹果的因素进行优化,通过含LFY基因的根癌农杆菌EHA105转化苹果叶片,获得了提早开花的转基因植株。结果表明:以OD600=0.3~0.5农杆菌菌液侵染叶片,25℃条件下共培养,共培养后的叶片延迟筛选3 d,在附加卡那霉素30 mg/L的再生培养基中选择培养,转化芽再生率可达6.97%。PCR检测初步证明目的LFY基因已整合到苹果基因组中。

关 键 词:苹果  农杆菌  遗传转化  LFY  GUS
文章编号:1000-1573(2006)03-0006-04
收稿时间:2005-12-12
修稿时间:2005年12月12

Study on the transformation of apple with LFY gene mediated by Agrobacterium tumefaciens
LIU Jing,SHAO Jian-zhu,XU Ji-zhong,LI Xiang-li,GUO Wen-wu.Study on the transformation of apple with LFY gene mediated by Agrobacterium tumefaciens[J].Journal of Agricultural University of Hebei,2006,29(3):6-9.
Authors:LIU Jing  SHAO Jian-zhu  XU Ji-zhong  LI Xiang-li  GUO Wen-wu
Abstract:Apple rootstock of M_(26) was transformed by Agrobacterium tumefaciens EHA105 carrying LFY gene.The factors affecting apple transformation were studied through GUS transient expression,such as different co-cultivation temperature,different bacterium density,et al.Leaf pieces were infected with Agrobacterium OD_(600)=0.3~0.5,then cocultivated at 25℃.After 3 d delayed selection the leaf pieces were incubated on regeneration medium containing 30 mg/L kanamycin.The regeneration rate of transformation buds reached 6.97%.It was proved by PCR analysis that the target LFY gene had been integrated into the genome of regenerated plants.
Keywords:apple  Agrobacterium tumefaciens  genetic transformation  LFY  GUS
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