• Title/Summary/Keyword: early globular embryo clusters

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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;Han, Seong-Soo;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.233-239
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    • 2003
  • 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.

The embryogenic competency and morphological changes during somatic embryogenesis in Iris pseudacorus

  • Kim, T.D.;Ahn, C.H.;Bae, K.H.;Choi, Y.E.
    • Plant Biotechnology Reports
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    • v.3 no.3
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    • pp.251-257
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    • 2009
  • Embryogenic callus was obtained from bulb segments of Iris pseudacorus on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or in combination with kinetin. When early globular somatic embryos were subcultured onto MS medium with $4.52{\mu}M$ 2,4-D, high frequency of somatic embryogenesis was obtained. Deprivation of 2,4-D was required for maturation. Mature somatic embryos had an elongated scutellum with a notch on the base of scutellum. Separation of embryos from embryo clusters was necessary to enhance the frequency of germination. Germination was stimulated by separation of embryos from embryo clusters and transfer onto fresh half-strength MS medium with 3% sucrose. After acclimation in artificial soil in greenhouse for 2 months, 96.4% of plantlets survived.

Agrobacterium-mediated transformation of Eleutherococcus senticosus with the squalene synthases gene derived from panax ginseng

  • Seo, Jin-Wook;Jeong, Jae-Hun;Han, Sung-Tai;Lee, Hak-Sung;Choi, Yong-Eui;Shin, Cha-Gyun
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.145.3-146
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    • 2003
  • Transgenic Eleutherococcus senticosus plants were prepared by introducing the genes for squalene synthase (SQS), hygromycin phosphotransferase (HPT) and green fluorescent Protein (GFP) through Agrobacterium-mediated transformation. The enzyme, SQS, represents a putative branch point in the isoprenoid pathway capable of diverting carbon flow specifically to the biosynthesis of phytosterol and oleanolic acid. The full SQS gene was isolated from P. ginseng roots. Early globular embryo clusters developed from embryogenic callus were used as the explant source. (omitted)

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Mass Propagation of Somatic Embryos and Plantlets of Aralia elata through Bioreactor Culture (생물반응기 배양을 통한 두릅나무(Aralia elata)의 체세포배 및 유식물체 대량증식)

  • Lee, Won-Seok;Choi, Eun-Gyung;Kim, Jae-Whune
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.219-223
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    • 2004
  • Embryogenic calli were induced from petioles of Aralia elata on MS solid medium supplemented with 1.0 mg/L 2,4-D. When embryogenic calli were transferred to MS liquid medium supplemented with 1.0 mg/L 2,4-D, embryogenic cells and embryogenic cell clusters were developed after 2 weeks of culture. Embryogenic cells were filtered through a 250 ${\mu}{\textrm}{m}$ sieve and the passed cells were proliferated and maintained in MS liquid medium supplemented with 1.0 mg/L 2,4-D. Embryogenic cell clusters entrapped on the sieve were transferred to 1/2 MS liquid medium without plant growth regulators, globular-shaped embryos were developed from embryogenic cell clusters after 2 weeks of culture. Numerous early stage somatic embryos could be developed to heart-shaped, torpedo-shaped, cotyledonary embryos and plantlets in 5 L bioreactor. Above results suggest that effective somatic embryo proliferation can be achieved via bioreactor culture systems in Aralia elata.