• Journal of Plant Biotechnology
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• 제30권3호
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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.

• Plant Biotechnology Reports
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• 제3권3호
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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.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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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)

• Journal of Plant Biotechnology
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• 제31권3호
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• pp.219-223
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• 2004

두릅나무의 엽병을 1.0 mg/L 2,4-D가 포함된 MS 고체배지에서 배발생캘러스를 유도하였다. 배발생세포와 배발생세포괴는 1.0 mg/L 2.4-D가 포함된 MS 액체배지에서 배발생캘러스를 2주간 현탁배양하여 대량으로 얻었다. 그물망을 통과한 배발생세포는 1.0 mg/L 2.4-D가 포함된 MS 액체배지에서 배양하면 배발생능이 소실되지 않고 지속적으로 유지 및 증식시킬 수 있었다. 그물망을 통과하지 못한 배발생세포괴를 식물생장조절물질이 첨가되지 않은 1/2 MS 액체배지에 옮겨 2주간 배양하면 구상형의 체세포배로 발달하였다. 구상형의 체세포배는 5 L의 bioreactor를 이용하여 배양하면 심장형, 어뢰형, 자엽형의 배와 유식물체로 발달하였다. Bioreactor 배양을 통해 두릅나무의 체세포배를 효과적으로 대량증식 시킬 수 있었다.