Agrobacterium-Mediated Transformation System for Large-Scale Producion of Transgenic Chinese Cabbage (Brassica rapa L. ssp. pekinensis) Plants for Insertional Mutagenesis
In order to better utilize insertional mutagenesis and functional genomics in Chinese cabbage, we have developed an improved transformation system that more efficiently produces a large number of transgenic plants. Hypocotyl explants were inoculated with Agrobacterium tumefaciens LBA4404. This strain harbors tagging vector pRCV2, which contains a hygromycin-resistance gene, an ampicillin resistance gene, and a bacterial replication origin within the T-DNA. Transformation efficiency was highest when the explants were first co-cultivated for 3 d in a medium supplemented with 5 mg L-1 acetosyringone, then transferred to a 0.8% agar selection medium containing 10 mg L-1 hygromycin. In addition, maintaining a low pH in the co-cultivation medium was critical to enhancing transformation frequency. A total of 3369 transgenic plants were obtained, with efficiencies ranging from 2.89% to 5.00%. Southern blot analysis and T1 progeny tests from 120 transgenic plants confirmed that the transgenes were stably inherited to the next generation. We also conducted plasmid rescue and inverse PCR with some transformants, based on their phenotype, to demonstrate the applicability of T-DNA tagging in Chinese cabbage. The tagged sequences were then analyzed.
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이 논문을 인용한 문헌 (4)
- Yu, Jae-Gyeong ; Lee, Gi-Ho ; Lim, Ki-Byung ; Hwang, Yoon-Jung ; Woo, Eun-Taek ; Kim, Jung-Sun ; Park, Beom-Seok ; Lee, Youn-Hyung ; Park, Young-Doo 2010. "Analysis of Mutant Chinese Cabbage Plants Using Gene Tagging System" 원예과학기술지 = Korean journal of horticultural science & technology, 28(3): 442~448
- Yu, Jae-Gyeong ; Park, Ji-Hyun ; Park, Young-Doo 2010. "Current status of Brassica rapa functional genome research in Korea" Journal of plant biotechnology = 식물생명공학회지, 37(2): 166~173
- 2010. "" Molecules and cells, 29(3): 267~275
- Yu, Jae-Gyeong ; Lee, Gi-Ho ; Park, Young-Doo 2012. "Comparison of RNA Interference-mediated Gene Silencing and T-DNA Integration Techniques for Gene Function Analysis in Chinese Cabbage" 원예과학기술지 = Korean journal of horticultural science & technology, 30(6): 734~742
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