农杆菌介导巨桉Eg5高效遗传转化

郭利军, 曾炳山, 刘英

doi:10.3724/SP.J.1259.2013.00087

植物学报 >

2013 , Vol. 48 >Issue 1: 87 - 93

DOI: https://doi.org/10.3724/SP.J.1259.2013.00087

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农杆菌介导巨桉Eg5高效遗传转化

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¹中国林业科学研究院热带林业研究所, 广州 510520;
²海南省农业科学院热带果树研究所, 海口 571100

收稿日期: 2012-06-11

修回日期: 2012-08-28

网络出版日期: 2012-11-01

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Agrobacterium-mediated High-efficient Transformation of Eucalyptus grandis Clone Eg5

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¹Research Institute of Tropical Forestry, Chinese Academy of Forestry, Guangzhou 510520, China

²Tropical Fruit Research Institute, Hainan Academy of Agricultural Sciences, Haikou 571100, China

Received date: 2012-06-11

Revised date: 2012-08-28

Online published: 2012-11-01

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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

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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

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.

Key words： Agrobacterium tumefaciens; Eucalyptus grandis; genetic transformation; selection strategies

参考文献

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