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| 农杆菌介导巨桉Eg5高效遗传转化 | |
| 引用本文： | 郭利军,曾炳山,刘英. 农杆菌介导巨桉Eg5高效遗传转化[J]. 植物学报, 2013, 48(1): 87-93. DOI: 10.3724/SP.J.1259.2013.00087 |
| 作者姓名： | 郭利军 曾炳山 刘英 |
| 作者单位： | 中国林业科学研究院热带林业研究所, 广州 510520; 海南省农业科学院热带果树研究所, 海口 571100 |
| 基金项目： | 高新技术研究发展计划(No.2011AA100202) |
| 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。 |
| 关 键 词： | 农杆菌 巨桉 遗传转化 筛选策略 |
| 收稿时间： | 2012-06-11 |
| 修稿时间： | 2012-08-28 |
Agrobacterium-mediated High-efficient Transformation of Eucalyptus grandis Clone Eg5 |
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| Lijun Guo,Bingshan Zeng,Ying Liu. Agrobacterium-mediated High-efficient Transformation of Eucalyptus grandis Clone Eg5[J]. Bulletin of Botany, 2013, 48(1): 87-93. DOI: 10.3724/SP.J.1259.2013.00087 | |
| Authors: | Lijun Guo Bingshan Zeng Ying Liu |
| Affiliation: | Research Institute of Tropical Forestry, Chinese Academy of Forestry, Guangzhou 510520, China Tropical Fruit Research Institute, Hainan Academy of Agricultural Sciences, Haikou 571100, China |
| Abstract: | 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 increased the selection efficiency to 15% and transformation efficiency was 0.26%. The plantlets were confirmed by GUS staining and PCR analysis. |
| Keywords: | Agrobacterium tumefaciens')" href=" #" >Agrobacterium tumefaciens Eucalyptus grandis')" href=" #" > Eucalyptus grandis genetic transformation selection strategies |
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