• Journal of Ginseng Research
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• v.38 no.3
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• pp.220-225
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• 2014

An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

• Korean Journal of Plant Resources
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• v.27 no.3
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• pp.242-248
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• 2014

This study was conducted to establish an efficient transformation system by using somatic embryogenesis in an important Korean native conifer, Korean fir (Abies koreana). Embryogenic masses were induced from mature zygotic embryos of the Korean fir on Schenk and Hildebrandt medium, which was supplemented with thidiazuron. For genetic transformation, the embryogenic masses were co-cultivated with a disarmed Agrobacterium tumefaciens strain C58/pMP90 containing the plasmid vector pBIV10 or LBA4404 containing the plasmid vector MP90. Both vectors contain the kanamycin resistance and beta-glucuronidase (GUS) reporter genes. A total of 48 lines of embryogenic masses were selected on mLV medium containing $50{\mu}g/mL$ of kanamycin after 4 weeks of culture, following 3 days of co-cultivation with A. tumefaciens strain C58/pMP90 carrying pBIV10 (none of the lines was cultivated with strain LBA4404 carrying MP90). Quantitative real-time PCR was performed, and high levels of GUS transcripts were observed in the 48 putative transgenic lines; however, the control (non-transgenic line) showed negative results. Results of histochemical staining showed that the expression of the GUS reporter gene was observed in somatic embryos that developed from the embryogenic masses of all 48 lines. Stably transformed cultures were successfully produced by co-cultivation with A. tumefaciens strain C58/pMP90 carrying pBIV10 in Korean fir. Here, we have reported an Agrobacterium-mediated gene transfer protocol via somatic embryogenesis that may be helpful in developing breeding and conservation strategies for the Korean fir.

• Korean Journal of Plant Tissue Culture
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• v.25 no.2
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• pp.115-118
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• 1998

This experiments were carried out to determine the optimum concentrations of plant growth regulators for the direct embryogenesis from the cotyledon culture of Paeonia lactiflora Pall. Zygotic embryos rescued from true seeds were developed abnormally in the medium containing ABA. But somatic embryogenesis from cotyledons of abnormal seedlings was induced more effectively. Also the somatic embryogenesis from cotyledons was promoted in the medium containing ABA. The frequency of embryogenesis was maximum(59.9%) from the cotyledons cultured on MS medium containing 0.5 mg/L ABA on which the frequency of somatic embryos with two cotyledons was 22.6%.

• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.343-356
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• 2010

Plant micropropagation techniques include bud cultures using apical or axillary buds, organogenesis through callus culture or adventitious bud induction, and somatic embryogenesis. In Korea Forest Research Institute (KFRI), the first tissue culture trial in woody plant was initiated from the bud culture of hybrid poplars (Populus alba x P. glandulosa) in 1978. Since then several mass propagation techniques have developed from conifer and hardwood species, resulting in allowing practical application to Poplars, Birches and some oak species. In addition, useful micropropagation and genetic resources conservation techniques were established in some rare and endangered tree species including Abeliophyllum distichum. Among various in vitro propagation techniques, somatic embryogenesis is known to be the most efficient plant regeneration system. Since the first somatic embryo induction was reported in Tilia amurensis by KFRI in 1986, various protocols for direct or indirect somatic embryogenesis systems have developed in conifer and hardwood species including Larix leptolepis, Pinus rigida x P. taeda F1, Kalopanax septemlobus and Liliodendron tulipifera, etc. However, most of these technologies have been developed using juvenile tissues, i.e. immature zygotic embryos or mature embryos. Therefore it has been difficult to directly application to tree breeding program due to their unproven genetic background. Recently remarkable progresses and new approaches have been achieved in mature tree somatic embryogenesis. In this article we reviewed several micropropagation techniques, which have been mainly developed by KFRI and recent international progresses.

• Plant Biotechnology Reports
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• v.2 no.1
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• pp.87-92
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• 2008

An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when cultured on halfstrength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly with increasing concentrations of 2,4-D up to $3mg\;l^{-1}$, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryoderived white friable callus were established using half-strength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of water-shield, which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could be useful for the mass propagation and transformation of selected elite lines.

• Korean Journal of Medicinal Crop Science
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• v.2 no.1
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• pp.60-66
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• 1994

This study was conducted to determine the optimum conditions of inducing callus, proliferating callus, forming somatic embryos, and regenerating plantlets via somatic embryogenesis, for the purpose of producing artificial seeds and substantially developing plant factory technologies that can be employed to all seasons production of Bupleurum plants. Callus was efficiently induced from leaf tissues at three leaf stage in the MS medium supplemented with 2, 4-D 2mg /1 and thidiazuron(TDZ) 0.lmg /1. Callus induction from leaf tissues at maturity was mostly effective in the mixture of 2,4- D 2mg /1 and TDZ 1.0mg /1 while that from flower bud tissues was fairly good in the MS medium containing 2,4-D 1 or 2mg /1.Callus was formed in 15 to 20 days after culture initiation in the MS media supplemented with 2, 4- D 1-2mg /1 and TDZ 0.l-1.0mg /1. Such hormones as kinetin 3mg /1, GA 1mg /1, and the mixture of GA 1mg /1 and TDZ 1mg /1 effected markedly to proliferate the callus cells.The optimum temperature and light intensity for callus culture were found to be $25^{\circ}C$ and 3000 Lux, respectively. Direct plant regeneration from cultured callus was fairly made on hormone-free MS or half-strength MS medium. Somatic embryogenesis was most frequently observed in hormone-free media:60 somatic embryos per 20ml in MS medium and 28 somatic embryos per 20ml in half -strength MS medium. There were three stages-globular, heart, and torpedo-in development of somatic embryos, among which globular stage was more frequently observed in MS medium rather than in half-strength MS medium. Somatic embryos induced from suspension culture fairly differentiated a number of shoots and roots on hormone-free and half-strength MS solid medium.

• Journal of Plant Biotechnology
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• v.40 no.3
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• pp.141-146
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• 2013

This study was conducted to examine suspended embryogenic cells growth with days of culture, effects of various kinds/concentrations of osmoticum for induction of somatic embryos (SEs), following somatic embryos germination or plantlet regeneration. The proliferation pattern of embryogenic cells in suspension culture is characterized by settled cells volume (SCV) increased with the duration of culture with marked the maxium of SCV (10.1 ml) in 18 days of culture, however the SCV of cells gradually decreased after that. In comparison of kinds/concentrations of osmoticum on somatic embryo induction, the highest induction number (352.3/g FW) of the SE was showed in 0.2 M sucrose, in addition, we also observed some effects with treatments of 0.2 M maltose (203.7) and 0.3 M maltose (193.7), respectively. However, no somatic embryos produced in treatments of 7.5% PEG plus 0.15 M sucrose or maltose. In comparison of germination efficiency of SEs which occurred from the treatments of various kinds/ concentrations of osmoticum, the highest induction frequency of cotyledon (25.2%) was obtained from SEs that produced 0.3 M maltose, however, the best occurrence rates of hypocotyl (39%), radicle (30.3%) and plantlet regeneration (3.5%) were observed from the 0.2 M sucrose treatment, respectively.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.171-171
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• 2003

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.63-63
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• 2004

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2000.11a
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• pp.27-33
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• 2000