Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Agrobacterium-mediated Transformation of Eleutherococcus sessiliflorus using Embryogenic Calli and the Regeneration of Plants

오갈피(Eleutherococcus sessiliflorus)의 배형성 세포를 이용한 고빈도 형질전환 및 재분화

  • Jeong, Jae-Hun (Korea Ginseng Institute, Chung-Ang University) ;
  • Han, Seong-Soo (College of Life Science Life Science and Natural Resources, Wonkwang University) ;
  • Choi, Yong-Eui (Korea Ginseng Institute, Chung-Ang University)
  • 정재훈 (중앙대학교 인삼산업연구센터) ;
  • 한성수 (원광대학교 생명자원과학대학) ;
  • 최용의 (중앙대학교 인삼산업연구센터)
  • Published : 2003.09.01
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Abstract

We have developed a reliable and high-frequency genetic transformation and regeneration system via somatic embryogensis of Eleutherococcus sessiliflorus. Embryogenic callus obtained from seed were co- cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring genes for intron-$\beta$-glucoronidase(GUS), kanamycin and hygromycin resistance. Following co-cultivation, two types of samples(fine embrogenic calli and early globular embryo clusters) were cultivated on Murashige and Skoog(MS) medium containing 1 mg/L2.4-D for 3day in dark. Transient expression of GUS gene was found to be higher in the early globular embryo clusters than in the embryogenic calli. Also, co-cultivated period affected expression of GUS gene; the best result was obtained when globular embryo clusters were co-cultivated with Agrobacterium for 3 days. Subsequently, this callus transferred to selective MS medium containing 1mg/L2.4-D, 50mg/L kanamycin or/and 30mg/L hygromycin and 300mg/L cefortaxime. These embryogenic calls were subcultured to the same selection medium at every 2 weeks intervals. Approximately 24.5% of the early globular embryos co-cultivated with Agrobacterium for 3days produced kanamycin or/and hygromycin-resistant calli. Transgenic somatic embryos were converted into plantlets in half strength MS medium supplemented with 3mg/L GA$_3$ kanamycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southem blot hybridization confirmed the incorporation of NPT II gene into the host genome.

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References

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