Transformation of RNAi Vector in Cucumber (Cucumis sativus) In Vitro by Agrobacterium tumefaciens-mediated Transfection

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  • Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China

Received date: 2013-01-26

  Revised date: 2013-04-12

  Online published: 2014-01-07

Abstract

The RNAi vector of the target gene was transfected into cucumber (Cucumis sativus) cotyledons and cotyledon nodes by Agrobacterium tumefaciens-mediated transfection. A total of 99 regeneration plants were hygromycin-resistant. PCR and Southern-blot analysis revealed that target gene was integrated into the genome of 10 regenerated plants. The transformation efficiency in the cucumber cotyledon and cotyledon node was significantly increased from 4.1% to 13.9% and 3.1% to 6.9%, respectively, when using different hygromycin concentrations in the selection cultivation. We have optimized the genetic transformation system with an RNA interference vector in cucumber and successfully obtained transgenic cucumber plants.

Cite this article

Ye Wang, Xingfang Gu, Shengping Zhang, Han Miao, Guohua Chen, Bingyan Xie . Transformation of RNAi Vector in Cucumber (Cucumis sativus) In Vitro by Agrobacterium tumefaciens-mediated Transfection[J]. Chinese Bulletin of Botany, 2014 , 49(2) : 183 -189 . DOI: 10.3724/SP.J.1259.2014.00183

References

韩欣 (2010). 根结线虫基因16D转化烟草和黄瓜的研究. 硕士学位论文. 长沙:湖南农业大学.
魏爱民,张文珠,杜胜利,韩毅科,张桂华,刘楠 (2008). 黄瓜花粉管通道法抗虫基因导入及卡那霉素抗性筛选. 华北农学报 23(6), 54-57.
魏爱民,张文珠,杜胜利,韩毅科,张桂华,张历 (2006). 影响农杆菌介导的黄瓜抗虫基因遗传转化体系的因素研究. 天津农业科学 12(3), 1-3.
于玉梅 (2008). 根结线虫16D10基因转化黄瓜的研究. 硕士学位论文. 长沙:湖南农业大学.
张若纬 (2010). 黄瓜离体再生和遗传转化体系的建立. 硕士学位论文. 北京:中国农业科学院研究生院.
张守杰 (2011). 黄瓜再生体系的建立及农杆菌介导的遗传转化. 硕士学位论文. 泰安:山东农业大学.
张文珠,巍爱民,杜胜利,韩毅科,张桂华,刘楠 (2009). 黄瓜农杆菌介导法与花粉管通道法转基因技术. 西北农业学报 18(1), 217-220.
朱妍妍 (2008). RNAi技术在黄瓜抗根结线虫遗传转化中的应用研究. 硕士学位论文. 泰安:山东农业大学.
Babinet C, Morello D, Renard JP. (1989). Transgenic mice. Genome, 31, 938-949.
Martin DIK, Whitlaw E. (1996). Vagaries of variegating trans-genes. BioEssays, 18(11), 919-923.
Eissenberg JC. (1989). Position effect variegation in Drosophila: towards a genetics of chromatin assembly. Bioessays, 11(1), 14-17.
Sambrook J, Fritsch EF, Maniatis T. (1989). Molecular cloning: A laboratory manual. 2nd ed. New York: Cold Spring Harbor Laboratory Press.
Selvaraj N, Kasthurirengan S, Vasudevan A, Manickavasagam M, Choi CW and Ganapathi A. (2010). Evaluation of green fluorescent protein as a reporter gene and phosphinothricin as the selective agent for achieving a higher recovery of transformants in cucumber (Cucumis sativus L. cv. Poinsett76) via Agrobacterium tumefaciens. In Vitro Cellular & Developmental Biology—Plant 46, 329–337.
Szwacka M, Malgorzata, Kielkiewicz (2007). Attractiveness of transgenic cucumber plants containing a gene encoding sweet-tasting protein to consumers and pests. Journal of Biotechnology 131(2), s37.
Vasudevan A, Selvaraj N, Ganapathi A. (2007). Agrobacterium-mediated Genetic Transformation in cucumber (Cucumis sativus L.). American Journal of Biotechnology and Biochemistry 3(1), 24-32.
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