• Korean Journal of Oriental Medicine
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• v.14 no.1
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• pp.113-116
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• 2008

Petiole explant of Glehnia littoralis Fr. Schmidt was in vitro cultured MS plant medium(DUCHEFA co.) supplemented with various plant growth regulators and examined to find out their optimum combination and concentration for plantlet regeneration. We investigated optimal condition for efficient plant regeneration through adventitious shoot formation on MS plant medium with various kinds of plant growth regulators. Embryogenic calli and adventitious shoot formation were greatly influenced by plant growth regulators. Embryogenic calli induction showed a good response on MS medium supplemented with NAA and BA than others. Especially, combination of 1.0 mg/L NAA and 0.5 mg/L BA on MS medium led to the greatest frequency in adventitious shoot. The results suggest that plant regulator selection be important factor to achieve an efficient regeneration.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.245-249
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• 2003

This study was carried out to establish a regeneration system from leaf explant of purple sweetpotato(Ipomoea batatas L.) The optimal concentrations of plant growth regulators for callus induction and shoot formation were determined. The optimal combination for callus formation was 1$\mu$M 2,4-D 5$\mu$M BM, and highest yield of embryogenic calli were observed on Murashige and Skoog basal medium containing 0.5$\mu$M 2,4-D under light condition after 4weeks of culture. Embryogenec callus was subcultured on medium supplemented with 5$\mu$M ABA for 4 days. Subsequently, regeneration of adventitious shoots occurred when these embryogenic calli were transferred onto medium with 3∼6$\mu$M gibberellic acid. Regenerated shoots were developed into normal plantlets.

• Korean Journal of Plant Resources
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• v.12 no.2
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• pp.107-119
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• 1999

In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.189-192
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• 2002

An efficient plant regeneration system of major cultivars of sweetpotato (Ipomoea batatas (L.) Lam.) in Korea via somatic embryogenesis was established. Embryogenic calli were formed from shoot apical meristems of sweetpotato cultivars when cultured on LS medium supplemented with 1 mg/L auxin (2,4-D, picloram, dicamba). Among three kinds of auxin, 1 mg/L 2,4-D showed the highest embryogenic calli induction rate. After 4 weeks of cultures on LS medium supplemented with 1 mg/L 2,4-D, embryogenic calli induction rates of Sinhwangmi, Zami, Yulmi, and White Star were 86%, 78%, 76%, and 80%, respectively. Upon transfer onto LS basal medium, most of somatic embryos developed into plantlets. Regenerated plantlets were transplanted to potting soil and grown to mature plants in a greenhouse.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.49 no.3
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• pp.237-242
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• 2004

A system for the production of transgenic plants has been developed for tall fescue (Festuca arundinacea Schreb.) via Agrobacterium-mediated transformation of mature seed-derived embryogenic callus. Seed-derived calli were infected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding the hygromycin phosphotransferase (HPT), neomycin phosphotransferase II (NPTII) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. Inclusion of $200\mu\textrm{M}$ acetosyringone (AS) in inoculation and co-culture media lead to a increase in stable transformation efficiency. Transformation efficiency was increased when embryogenic calli were co-cultured for 5 days on the co-culture medium. The highest transformation efficiency was obtained when embryogenic calli were inoculated with Agyobacterium in the presence of 0.1% Tween20 and $200\mu\textrm{M}$ AS. Hygromycin resistant calli were developed into complete plants via somatic embryogenesis. GUS histochemical assay and Southern blot analysis of transgenic plants demonstrated that transgenes were successfully integrated into the genome of tall fescue.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.263-268
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• 2005

Porcine epidemic diarrhea virus (PEDV) causes acute enteritis in pigs of all ages and is often fatal for neonates. In order to develop sweetpotato plants expressing PEDV antigen, we constructed the vector expressing spike gene of PEDV under the control of sweetpotato sporamin promoter or constitutive CaMV 35S promoter. The spike protein region of PEDV was synthesized by PCR and linked to each promoter, Transgenic sweetpotato [Ipomoea batatas (L.) Lam. cv. Yulmi] plants were developed from embryogenic calli following Agrobacterium tumefaciens-mediated transformation. The co-cultured embryogenic calli transferred to selective MS medium containing 1 mg/L 2,4-D, 100 mg/L kanamycin, and 400 mg/L claforan. These embryogenic calli were subcultured to the same selection medium at 3 weeks interval. Kanamycin-resistant calli transferred to hormone-free MS medium with kanamycin gave rise to somatic embryos and then converted into plantlets in the same medium. Southern blot analysis confirmed that the spike gene of PEDV was inserted into the genome of the sweetpotato plants. RT-PCR revealed that the spike gene of PEDV was highly expressed in transgenic sweetpotato plants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.19 no.3
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• pp.273-280
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• 1999

Hypocotyl explants of Medicago saliva cv. Vernal were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of growth regulators. After six weeks of culture, somatic embryos were formed from calli on MS medium containing $4mg/{\ell}$ 2,4-D and $0.1mg/{\ell}$ kinetin, or $4mg/{\ell}$ 2,4-D and $0.5mg/{\ell}$ kinetin. The mature somatic embryos were developed to plantlets when subcultured on MS basal medium. In order to obtain secondary somatic embryogenic calli, cotyledon of regenerated plantlets were cultured on a callus induction medium. Embryogenic calli were formed on MS medium containing $4mg/{\ell}$ 2,4-D alone. For induction and development of secondary somatic embryogenesis, the embryogenic calli were transferred to MS basal medium containing either 2,4-D or NAA. Multi-secondary somatic embryogenesis was the most effective on MS basal medium with $0.1mg/{\ell}$ 2,4-D. The rate of secondary somatic embryo formation of regenerated plants was 18 times higher than that of seed grown plants. The mature secondary somatic embryo were germinated into plantlets on MS basal medium after six weeks of culture.

• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.235-242
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• 2004

A system for the production of transgenic plants has been developed for Italian ryegrass(Lolium mult리orum Lam.) via Agrobacterium-mediated transformation of embryogenic callus. Mature seed-derived calli were infected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding the hygromycin phosphotransferase(HPT), neomycin phosphotransferase II (NPTII) and intron-oontaining $\beta$g1ucuronidase( intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. Inclusion of 200${\mu}M$ acetosyringone(AS) in inoculation and co-cultivation media lead to a significant increase in stable transformation efficiency. Increasing Agrobacterium cell density up to 1.0 in $OD_{600}$ during infection increased transfonnation efficiency of embryogenic calli. The highest transfonnation efficiency was obtained when embryogenic calli were incoulated with Agrobacterium in the presence of 0.1% Tween20 and 200${\mu}M$ AS. Hygromycin resistant calli were developed into complete plants via somatic embryogenesis. GUS histochemical assay and PCR analysis of transgenic plants demonstrated that transgenes were integrated into the genome of Italian ryegrass.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.388-395
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• 2015

Somatic embryogenesis is as an excellent technology for potential use in plant mass production, germplasm conservation, or genetic engineering. We examined the effect of cold storage using 3 embryogenic callus lines with different levels of embryogenesis competence derived from immature zygotic embryo cultures of Kalopanax setemlobus. Somatic embryo induction, germination and plant conversion were evaluated after 1, 3 and 6 months storage at $4^{\circ}C$ in the dark. Most cold-stored embryogenic calli formed somatic embryos normally even after 6 months; however, the induction rate was gradually decreased by increasing the storage period. The most competent line tended to show a slight decline in somatic embryo induction rate, as compared with other lines after cold storage. In general, cold storage resulted in reduced somatic embryo germination and plant regeneration, although 93% somatic embryo germination and 91% plant conversion were achieved regardless of the storage period. Cold storage led to cell browning and degradation. Additionally, the cell structures were confirmed by the aceto-carmine and evans blue dye evaluation. Collectively, our results showed that embryogenic callus of K. septemlobus could be preserved at $4^{\circ}C$ without subculture for 6 months, and suggested the need for storage of relatively more competent embryogenic calli lines to support somatic embryo induction.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.77-81
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• 1995

When cultured on MS medium supplemented with 0.5 mg/L NAA alone or 1.0 mg/L 2,4-D and 0.5 mg/L BA, immature zygotic embryos of Stewartia koreana formed embryogenic calli and somatic embryos. In investigate effect of sucrose concentration on somatic embryo development, embryogenic calli were transferred to MS basal medium containing 1.5,3, 6 or 9% sucrose. The greatest frequency of somatic embryos was obtained on medium containing 6% sucrose. However addition of 1.5 or 9% sucrose to medium inhibited somatic embryo germination and development into normal plantlet After 5 weeks of hardening culture on medium containing 6% sucrose, somatic embryos were transferred to half strangth MS medium supplemented with 0.1% charcol, wherein these embryo developed into the normal plantlets.