摘要
为建立和优化根癌农杆菌介导的大麦遗传转化体系,以大麦幼胚为转化受体,研究了根癌农杆菌介导法转化大麦的主要影响因素。结果表明,以gus基因瞬时表达率、抗性愈伤组织诱导率为转化指标,筛选出了对大麦幼胚感染力比较强的农杆菌菌株EHA105,以及高敏感受体基因型甘啤4号和秀81-47等。实验结果还表明,预培养1 d的大麦幼胚具有较高的瞬时表达率(91%),是较好的转化受体。菌液侵染浓度OD600=0.5,侵染时间为30 min时转化效率最高。在此基础上,侵染后幼胚与农杆菌采用液体共培养方式可明显提高转化效率,gus基因瞬时表达率最高可达90%。在优化条件下,转化约30 d后抗选择剂潮霉素(Hyg)的愈伤组织占受体组织总数的34.2%。
To establish and optimize barley transformation system by Agrobacterium-mediated,immature embryos were used as target materials.Important factors that influence Agrobacterium-mediated transformation of barley were investigated.The results showed that,using transient GUS expression,frequency of resistant calli as a criteria,one highly infectious agrobacterium tumefaciens strains（EHA105）to barley and some highly sensitive barley acceptors（Ganpi4 and Xiu81-47） were obtained.The barley immature embryos were pre-cultured for 1 day,and using the Cell density of OD600=0.5 and 30 minutes to inoculate immature embryos,which was optimal conditions for Agrobacterium-mediated transformation of barley immature embryos.Under these optimal conditions,hygromycin resistant calli was 34.2% of of total explants infected after about 30 days.
出处
《大麦与谷类科学》
2010年第2期1-6,共6页
Barley and Cereal Sciences
基金
甘肃省农牧厅生物技术专项"啤酒大麦籽粒蛋白质表达的遗传调控及种质创新"(GNSW-2006-05)
关键词
大麦
幼胚
根癌农杆菌
遗传转化
Barley
Immature embryo
Aarobacterium tumefaciens
Genetic transformation
