文献类型：期刊文献

中文题名：农杆菌介导巨桉Eg5高效遗传转化

英文题名：Agrobacterium-mediated High-efficient Transformation of Eucalyptus grandis Clone Eg5

作者：郭利军[1,2] 曾炳山[1] 刘英[1]

第一作者：郭利军

机构：[1]中国林业科学研究院热带林业研究所;[2]海南省农业科学院热带果树研究所

年份：2013

卷号：48

期号：1

起止页码：87-93

中文期刊名：植物学报

外文期刊名：Chinese Bulletin of Botany

收录：CSTPCD;;北大核心:【北大核心2011】；CSCD:【CSCD2013_2014】；

基金：高新技术研究发展计划(No.2011AA100202)

语种：中文

中文关键词：农杆菌;巨桉;遗传转化;筛选策略

外文关键词：Agrobacterium tumefaciens, Eucalyptus grandis, genetic transformation, selection strategies

分类号：Q943.2

摘要：以巨桉(Eucalyptus grandis)无性系Eg5叶片为外植体,探讨了农杆菌(Agrobacterium tumefaciens)侵染时间、共培养pH值和共培养时间对瞬时转化效率的影响,分析了不同筛选策略对遗传转化植株筛选效果的影响。结果表明,外植体侵染45分钟,共培养pH值为5.8,共培养3天所得到的瞬时转化效率最高;逐步提高卡那霉素(Km)浓度筛选转基因植株有效,筛选率达到15%,转化率达到0.26%。经过GUS染色分析和PCR检测,证实为转基因植株。
In this study, we used Eucalyptus grandis clone Eg5 leaves as explants to study the influence of infection time, co-culture pH values and co-culture time on transient expression efficiency. We analyzed the effects of different selection strategies on transgenic plants. Infection time of 45 min, co-culture pH of 5.8, and co-culture time of 3 days achieved the best transient expression efficiency. Screening transgenic plantlets by gradually improving kanamycin concentration in- creased the selection efficiency to 15% and transformation efficiency was 0.26%. The planUets were confirmed by GUS staining and PCR analysis.