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Improvement of Transformation Efficiencies using Agrobacterium-Mediated Transformation of Korean Rice
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  • Improvement of Transformation Efficiencies using Agrobacterium-Mediated Transformation of Korean Rice
  • Improvement of Transformation Efficiencies using Agrobacterium-Mediated Transformation of Korean Rice
저자명
Cho. Joon-Hyeong,Lee. Jang-Yong,Kim. Yong-Wook,Lee. Myoung-Hoon,Park. Seong-Ho
간행물명
Korean journal of crop science
권/호정보
2004년|49권 1호|pp.61-68 (8 pages)
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한국작물학회
파일정보
정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

A reproducible transformation system via optimized regeneration media for Korean rice cultivars was established using Agrobacterium tumefeciens LBA4404 (pSBM-PPGN; gusA and bar). Although japonica rice genotypes were easier to produce transgenic plants compared to Tongil type cultivars, transformation efficiencies were not always correlated with regeneration efficiencies of non-transgenic callus on the control medium. Regeneration efficiencies of Donganbyeo, Ilmibyeo, and Manchubyeo were over 50% in non-transgenic control, however, transformation efficiencies were significantly low when only sucrose was added to the media as a carbon source. However, the medium, MSRK5SS-Pr (or MSRK5SM-Pr), that contains $5 extrm{mgL}^{-1}$ kinetin, $0.5 extrm{mgL}^{-1}$ NAA, 2 % sucrose (or maltose), 3% sorbitol, and $500 extrm{mgL}^{-1}$ proline, was the most efficient not only for regeneration of non-transgenic callus but also for regeneration of transgenic callus in the presence of L-phosphinotricin (PPT). Average transformation efficiencies of 16 Korean rice cultivars were significantly enhanced by using the optimized medium from 1.5% to 5.8% in independent callus lines and from 2.9% to 19.4% in tromsgenic plants obained. Approximately 98.9% (876 out of 885) transgenic plants obtained on optimized media showed basta resistance. Stable integration, inheritance and expression of gusA and bar genes were continued by GUS assay and PCR and Southern analysis of the bar gene. With Pst1 digestion of genomic DNA of transgenic plants, one to five copies of T-DNA segment were observed; however, 76% (19 out of 25 transgenic plants) has low copy number of T-DNA. The transformants obtained from one callus line showed the same copy numbers with the same fractionized band patterns.