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

doi:10.3724/SP.J.1259.2014.00183

Abstract

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.

Key words: cucumber, RNAi vector, cotyledon, cotyledon node, transformation

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.

Add to citation manager EndNote|Ris|BibTeX

URL: https://www.chinbullbotany.com/EN/10.3724/SP.J.1259.2014.00183

https://www.chinbullbotany.com/EN/Y2014/V49/I2/183

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.

[1]	Jingjing Li, Yanfei Li, Anqi Wang, Jiaying Wang, Chengyan Deng, Min Lu, Jianying Ma, Silan Dai. Establishment of Regeneration and Genetic Transformation System for Chrysanthemum Cultivar ‘Wandai Fengguang’ [J]. Chinese Bulletin of Botany, 2025, 60(4): 1-0.
[2]	Zeng Wendan, Yan Huabing, Wu Zhengdan, Shang Xiaohong, Cao Sheng, Lu Liuying, Xiao Liang, Shi Pingli, Cheng Dong, Long Ziyuan, Li Jieyu. Agrobacterium rhizogenes-mediated Transformation System of Pueraria lobata Hairy Roots [J]. Chinese Bulletin of Botany, 2025, 60(3): 425-434.
[3]	Xinyi He, Yumei Pan, Yan Zhu, Jiayi Chen, Sirong Zhang, Naili Zhang. Impact of ectomycorrhizal tree dominance and species richness on soil nitrogen turnover in a warm temperate forest [J]. Biodiv Sci, 2024, 32(9): 24173-.
[4]	Yuchen Li, Haixia Zhao, Xiping Jiang, Xintian Huang, Yaling Liu, Zhenying Wu, Yan Zhao, Chunxiang Fu. Establishment of Agrobacterium-mediated Transformation System for Agropyron mongolicum [J]. Chinese Bulletin of Botany, 2024, 59(4): 600-612.
[5]	Yu Xiaomin, Wang Yaqin, Liu Yuhan, Yi Qingping, Cheng Wenhan, Zhu Yu, Duan Feng, Zhang Lixue, He Yanhong. Establishment of Agrobacterium tumefaciens-mediated Genetic Transformation System of Marigold (Tagetes erecta) [J]. Chinese Bulletin of Botany, 2023, 58(5): 760-769.
[6]	YANG Yuan-He, ZHANG Dian-Ye, WEI Bin, LIU Yang, FENG Xue-Hui, MAO Chao, XU Wei-Jie, HE Mei, WANG Lu, ZHENG Zhi-Hu, WANG Yuan-Yuan, CHEN Lei-Yi, PENG Yun-Feng. Nonlinear responses of community diversity, carbon and nitrogen cycles of grassland ecosystems to external nitrogen input [J]. Chin J Plant Ecol, 2023, 47(1): 1-24.
[7]	Qiong Zhai, Rongqin Chen, Xiaohua Liang, Chuchun Zeng, Bo Hu, Ling Li, Xiaoyun Li. Establishment and Application of a Rapid Genetic Transformation Method for Peanut [J]. Chinese Bulletin of Botany, 2022, 57(3): 327-339.
[8]	Binqi Li, Jiahui Yan, Hao Li, Wei Xin, Yunhe Tian, Zhenbiao Yang, Wenxin Tang. Changes of Small GTPases Activity During Cucumber Tendril Winding [J]. Chinese Bulletin of Botany, 2022, 57(3): 299-307.
[9]	XIE Huan, ZHANG Qiu-Fang, ZENG Quan-Xin, ZHOU Jia-Cong, MA Ya-Pei, WU Yue, LIU Yuan-Yuan, LIN Hui-Ying, YIN Yun-Feng, CHEN Yue-Min. Effects of nitrogen addition on phosphorus transformation and decomposition fungi in seedling stage of Cunninghamia lanceolata [J]. Chin J Plant Ecol, 2022, 46(2): 220-231.
[10]	Lan Yang, Ya Liu, Yang Xiang, Xiujuan Sun, Jingwei Yan, Aying Zhang. Establishment and Optimization of a Shoot Tip-based Genetic Transformation System for Foxtail Millet [J]. Chinese Bulletin of Botany, 2021, 56(1): 71-79.
[11]	Xin Liu, Xiaoying Rong, Yuanming Zhang. Biocrusts impact niche separation of ammonia oxidizing microorganisms in the Gurbantunggut Desert, northwestern China [J]. Biodiv Sci, 2021, 29(1): 43-52.
[12]	Pengfei Du, Yu Wang, Yingping Cao, Song Yang, Zhichao Sun, Decai Mao, Jiajun Yan, Daxu Li, Meizhen Sun, Chunxiang Fu, Shiqie Bai. Establishment of Biolistic Mediated Transformation System for Elymus sibiricus [J]. Chinese Bulletin of Botany, 2021, 56(1): 62-70.
[13]	Yue Xu,Yingping Cao,Yu Wang,Chunxiang Fu,Shaojun Dai. Agrobacterium rhizogenes-mediated Transformation System of Spinacia oleracea [J]. Chinese Bulletin of Botany, 2019, 54(4): 515-521.
[14]	Jia Guo,Yansu Li,Chaoxing He,Yan Yan,Xianchang Yu. Establishing a High-efficiency Regeneration System in Pumpkin (Cucurbita moschata) [J]. Chinese Bulletin of Botany, 2019, 54(4): 539-546.
[15]	Li Zhu, Qian Qian. Astaxanthin Functional Rice: New Idea of Biofortification, New Perspectives for High-quality Rice Breeding [J]. Chinese Bulletin of Botany, 2019, 54(1): 4-8.