• Journal of Plant Biology
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• v.36 no.3
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• pp.211-218
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• 1993

Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

• Korean Journal of Plant Resources
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• v.24 no.3
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• pp.280-285
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• 2011

In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

• Journal of Plant Biology
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• v.34 no.1
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• pp.19-23
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• 1991

Culture of embryogenic callus and suspension were induced from rice seeds in MS2.5 medium. In hormone free N6 medium, whole plantlets were regenerated from embryogenic callus. We observed cell division and reformation of embryogenic callus on culture of protoplast isolated from embryogenic cell suspensions. In addition, we studied the influencing factors on viability of protoplast treated with electroporation. Viability was decreased according to the increase of voltage and capacitance during electroporation. An optimal level of viability was obtained after treatment with $200-300\;V/1180\;\mu\textrm{F}$ in HEM buffer at $4^{\circ}C$..

• Journal of Plant Biotechnology
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• v.3 no.2
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• pp.101-106
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• 2001

In order to establish efficient plant regeneration from embryogenic suspension cultures of soybean, Glycine max L, we examined the effects of auxin type and concentration, cytokinin type and concentration, and amino acid type and concentration on the growth of embryogenic clumps from induced callus, and the effect of desiccation of mature somatic embryos obtained from these clumps on the frequency of somatic embryo germination. Embryogenic callus was induced from the edge of the cotyledons cultured on MS medium containing 6% sucrose, 40 mg/L 2,4-D, 0.2% gelrite and pH 5.7. The growth of embryogenic clumps was best in early staged, embryogenic callus that was placed in suspension culture of MS medium containing 5 mg/L 2,4-D and 0.5 mg/L asparagine. Single somatic embryos were isolated from the clumps and plated on the same medium for maturation. When the mature single somatic embryos were desiccated for 96 h, somatic embryo germination came up to approximately 90%. The plantlets germinated after embryos desiccation for 2 weeks were transfered to MS medium containing 3% sucrose,0.2% gelrite and pH 5.7.

• Journal of Plant Biology
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• v.31 no.1
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• pp.41-49
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• 1988

Several cultivars of rice were examined for induction of embryogenic callus on a medium containing MS salts, vitamins and 2, 4-D under darkness. Embryogenic callus was obtained from cultivar Cheonma with high ratio and embryo-like structures were formed from the callus on a medium with or without reduced 2, 4-D. Somatic embryoids with a plumule and radicle axis surrounded by a scutellum were observed. These embryoids germinated and produced plantlets in 30 days on the same medium. Protoplasts isolated from an embryogenic cell suspension culture derived from embryogenic callus were cultured either in liquid or in agar medium and protoplast derived cell colonies were obtained in 3-4 weeks.

• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.289-291
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• 1999

Hypocotyl explants from 7 days old seedlings of one $F_1$ hybrid cultivar and two pure lines of cucumber formed embryogenic calli at frequencies of up to 8% when cultured on Murashige and Skoog medium (MS) supplemented with 1 mg/L 2,4-D for 3 weeks. Embryogenic calli gave rise to somatic embryos. When slices of somatic embryos were cultured on the same medium for 4 weeks, they formed embryogenic calli. Embryogenic cell suspension cultures were established with embryogenic calli in MS liquid medium with 1 mg/L 2,4-D. Embryogenic potential of cell suspension cultures was maintained by subculturing every seven days. When the level of 2,4-D in the medium was lowered to 0.2 mg/L by diluting with liquid MS basal medium, embryogenic cell suspension cultures underwent development into numerous somatic embryos. When plated onto MS basal medium, over 95% of somatic embryos developed into plantlets. Plantlets were transplanted to potting soil and grown to maturity.

• Journal of Plant Biology
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• v.33 no.4
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• pp.259-269
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• 1990

Changes of peroxidase activity, polyamine content and ethylene production during somatic embryogenesis in suspension-cultured carrot (Daucus carota L.) cells were investigated. As compared with nonembyrogenic cells and their medium, embryogenic cells and their medium were characterized by higher levels of peroxidase at all times of culture period. Peroxidase in embryogenic cells showed higher oxidation activity of IAA than in nonembryogenic cells at the torpedo stage, but the IAA oxidation activity of peroxidase released into embryogenic medium was lower than that of peroxidase released into nonembryogenic medium. Peroxidase patterns of embryogenic and nonembryogenic cells showed three cathodic bands, and one anodic band, while peroxidase patterns released into embryogenic and nonembryogenic media did not show any anodic bands and the isoelectric points of cathodic peroxidase were pH 7.7, 7.5 and 6.6. Compared with nonembryogenic cells, polyamine content in embryogenic cells was increased by 15% at the torpedo stage, but polyamine ratio was constant, and ethylene production was extremely low at all times of culture period. Therefore, it is suggested that the peroxidase in embryogenic cells is correlated with embryogenesis by regulating hormone ratios through IAA oxidation, while the peroxidase isozyme patterns may be used as a biochemical marker of embryogenesis. The increase of polyamine content and the decrease of ethylene production suggest an interaction between polyamine and ethlyene during embryogenesis.

• Asian Journal of Turfgrass Science
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• v.23 no.2
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• pp.345-352
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• 2009

Zoysiagrass (Zoysia japonica Steud) is a warm season turfgrass species widely used for sports field and golf courses. Many cultivars are propagated through vegetative methods. This study was conducted to develop an optimum culture medium and culture conditions for embryogenic callus induction and plant regeneration, and to establish a cell suspension culture system for use in zoysiagrass breeding and propagation. The results indicated that adding $Cu^{++}$ at 2.5 mg $L^{-1}$ to the induction medium was optimum for callus induction. Increasing the numbers of sub-culture cycles improved the quality of calli. The optimum dosage for cell suspension culture ranged from 2.5 to 10 mL. The embryogenic callus suspension used in this study had a plant regeneration rate of 58%.

• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.181-187
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• 2007

Orostachys japonicus A. Berger is a Perennial herbaceous plant which has been traditionally used as an anti-inflammatory agent to treat hepatitis and as an anticancer agent. The objective of this study was 1) to establish and proliferate in vitro plant of O. japonicus 2) to induce indirect somatic embryogenesis from O. japonicus. General calli and embryogenic calli in all ranges of 2,4-D and BA combination, were induced and were best at 22% (embryogenic cell) in 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. Embryogenic cell line was maintained by subculture at 2 week intervals and transferred to solid and liquid medium for embryo formation. In solid medium culture, globular and heart shaped embryos were observed in MS medium containing 5.0 mg/L 2,4-D and 0.5 mg/L BA combination. The number of embryos was 6.5 per 0.5 g cell, and then the immature embryos transferred to MS basal medium for embryo development. In a suspension culture of embryogenic cells, globular and heart shaped embryos were emerged in MS medium supplemented with 3.0 mg/L 2,4-D and 0.3 mg/L BA combination after 10 days of incubation. The embryo formation rate was about 33% by suspension culture. The ratio of embryo germination was 60.9%, on the other side, the root formation rate was 74.3% in 1/2 MS continuously.

• Korean Journal of Medicinal Crop Science
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• v.1 no.2
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• pp.184-190
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• 1993

This study was carried out to investigate the appropriate medium and constitutionof growth regulators for somatic embryogenesis for development of rapid mass propagation system via somatic embrygenesis in Rehmennia glutinosa. Embryogenic callus formation from leaf explant was more effective when 4mg / l BA with 0.5mg / l NAA than that of treated with only auxins or cytokinins. LS medium was suitable for embryogenic callus formation. LS medium with 4mg / l BA with 0.5mg / l NAA was effective for the maintenance and proliferation of embryogenic callus. In suspension culture, addition of 1mg / l BA to LS medium was proper for somatic embryogenesis. The highest rate of shoot developement form cotyledon stage embryo was obtained in 1/2 LS medium and plantlet survived by 75% after transplanted to the soil. after 4 weeks.