农杆菌介导巨桉Eg5高效遗传转化

doi:10.3724/SP.J.1259.2013.00087

植物学报

农杆菌介导巨桉Eg5高效遗传转化

郭利军^1,2, 曾炳山^1*, 刘英¹

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 出版日期:2013-01-01 发布日期:2012-11-01
通讯作者: 曾炳山

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

Lijun Guo^1,2, Bingshan Zeng^1*, Ying Liu¹

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:2012-06-11 Revised:2012-08-28 Online:2013-01-01 Published:2012-11-01
Contact: Bingshan Zeng

摘要/Abstract

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。

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.

中图分类号:

S792.39 Q943.2

郭利军, 曾炳山, 刘英. 农杆菌介导巨桉Eg5高效遗传转化. 植物学报, DOI: 10.3724/SP.J.1259.2013.00087.

Lijun Guo, Bingshan Zeng, Ying Liu. Agrobacterium-mediated High-efficient Transformation of Eucalyptus grandis Clone Eg5. Chinese Bulletin of Botany, DOI: 10.3724/SP.J.1259.2013.00087.

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参考文献

参考文献:
Aggarwal, D., A. Kumar,M. Sudhakara Reddy (2011). Agrobacterium tumefaciens mediated genetic transformation of selected elite clone(s) of Eucalyptus tereticornis. Acta Physiologiae Plantarum, 33, 1603.
Alcantara, G. B. D., J. C. Bespalhok Filho,M. Quoirin (2011). Organogenesis and transient genetic transformation of the hybrid Eucalyptus grandis × Eucalyptus urophylla . Scientia Agricola, 68, 246-251.
Cervera, M., J. A. Pina, J. Juarez, L. Navarro,L. Pena (1998). Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration. Plant Cell Reports, 18, 271-278.
Da Silva, A. L. L., Y. de Oliveira,J. Da Luz Costa (2011). Preliminary results for genetic transformation of shoot tip of Eucalyptus saligna Sm. via Agrobacterium tumefaciens. Journal of Biotechnology and Biodiversity, 2, 1-6.
Fillatti, J. A. J., J. Sellmer, B. McCown, B. Haissig,L. Comai (1987). Agrobacterium mediated transformation and regeneration of Populus. Molecular and General Genetics MGG, 206, 192-199.
González, E. R., A. de Andrade, A. L. Bertolo, G. C. Lacerda, R. T. Carneiro, V. A. P. Defávari, M. T. V. Labate,C. A. Labate (2002). Production of transgenic Eucalyptus grandis x E. urophylla using the sonication-assisted Agrobacterium transformation (SAAT) system. Functional plant biology, 29, 97-102.
Ho, C. K., S. H. Chang, J. Y. Tsay, C. J. Tsai, V. L. Chiang,Z. Z. Chen (1998). Agrobacterium tumefaciens-mediated transformation of Eucalyptus camaldulensis and production of transgenic plants. Plant Cell Reports, 17, 675-680.
Jefferson, R. A., T. A. Kavanagh,M. W. Bevan (1987). GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The EMBO journal, 6, 3901-3907.
J. L. Yao ,K. Lin-Wang. (2004). Eucalyptus transformation method. USA patent. PCT/NZ2004/000242. 2005-04-14.
Joyce, P., M. Kuwahata, N. Turner,P. Lakshmanan (2010). Selection system and co-cultivation medium are important determinants of Agrobacterium-mediated transformation of sugarcane. Plant Cell Reports, 29, 173-183.
Kajita, S., K. Osakabe, Y. Katayama, S. Kawai, Y. Matsumoto, K. Hata,N. Morohoshi (1994). Agrobacterium-mediated transformation of poplar using a disarmed binary vector and the overexpression of a specific member of a family of poplar peroxidase genes in transgenic poplar cell. Plant Science, 103, 231-239.
Mohri, T., N. Yamamoto,K. Shinohara (1996). Agrobacterium-mediated transformation of lombardy poplar (Populus nigra L. var. italica Koehne) using stem segments. Journal of Forest Research, 1, 13-16.
Mullins, K. V., D. J. Llewellyn, V. J. Hartney, S. Strauss,E. S. Dennis (1997). Regeneration and transformation of Eucalyptus camaldulensis. Plant Cell Reports, 16, 787-791.
Oliveira, Y., L. Adamuchio, J. Degenhardt-Goldbach, I. Gerhardt, J. Bespalhok, R. Dibax,M. Quoirin. 2011. Use of kanamycin for selection of Eucalyptus saligna genetically transformed plants., 147-148.: BioMed Central Ltd.
Padilla, I.,L. Burgos (2010). Aminoglycoside antibiotics: structure, functions and effects on in vitro plant culture and genetic transformation protocols. Plant Cell Reports, 29, 1203-1213.
Petri, C., S. López-Noguera, N. Alburquerque, J. Egea,L. Burgos (2008). An antibiotic-based selection strategy to regenerate transformed plants from apricot leaves with high efficiency. Plant Science, 175, 777-783.
Prakash, M.,K. Gurumurthi (2009). Genetic transformation and regeneration of transgenic plants from precultured cotyledon and hypocotyl explants of Eucalyptus tereticornis Sm. using Agrobacterium tumefaciens. In Vitro Cellular & Developmental Biology - Plant, 45, 429-434.
Spokevicius, A. V., K. Van Beveren, M. A. Leitch,G. Bossinger (2005). Agrobacterium-mediated in vitro transformation of wood-producing stem segments in eucalypts. Plant cell reports, 23, 617-624.
Thakur, A. K., S. Sharma,D. K. Srivastava (2005). Plant regeneration and genetic transformation studies in petiole tissue of Himalayan poplar (Populus ciliata Wall.). Current Science, 89, 664-668.
Wei, T. (2000). Genetic transformation of loblolly pine using mature zygotic embryo expiants by Agrobacterium tumefaciens. Journal of Forestry Research, 11, 215-222.
Wei, T. (2001). Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.). Cell Research, 11, 237-243.
樊军锋 (2002). mtlD/gutD 双价耐盐基因转化杨树, 猕猴桃的研究. 博士论文. 杨凌:西北农林科技大学. pp.57-59.
方中达 (1979). 植病研究方法. 北京:农业出版社. pp. 1-12.
郭利军, 曾炳山, 刘英, 李湘阳,裘珍飞 (2012). 巨桉无性系Eg5的卡那霉素和头孢霉素敏感性研究. 中南林业科技大学学报, 32, 75-80.
姜清彬 (2011). 细枝木麻黄再生体系及农杆菌介导遗传转化研究. 博士论文. 北京:中国林业科学研究院. pp.73-74.
林平 (2001) 永安桉树造林及其冻害情况调查. 桉树科技, 17, 15-20.
祁述雄 (2002). 中国桉树 (第2版). 北京:中国林业出版社. pp. 228-230.
曾炳山, 张剑, 李建娟, 刘英, 陈李花,范春节 (2010). 桉树人工林冰雪灾害恢复的调查. 中南林业科技大学学报, 30, 69-74.

农杆菌介导巨桉Eg5高效遗传转化

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

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