• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.169-174
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• 2005

The optimal culture conditions were studied for plant regeneration from mesophyll protoplasts of Solanum sisymbriifolium. Axenic seedlings of S. sisymbriifolium were used as a explant for protoplast culture. Many viable protoplasts were isolated by incubating leaf slices in an enzyme solution containing 0.25% Meicerase and 0.05% Macerozyme for 16 hr at $25^{\circ}C$ without shaking. Protoplast density of $5.0{\times}10^4\;ml^{-1}$ in Kao medium containing 5.0 mg/L NAA, 1.0 mg/L 2,4-D and 1.0 mg/L BA was optimal for colony formation. Most colonies were formed when protoplasts were cultured at $25^{\circ}C$ after initial culture at $30^{\circ}C$ for one week. On the MS agar medium with 1.0 mg/L zeatin, 38.4% of protoplast-derived calli differentiated shoots. These shoots rooted on 1/2MS medium with 5.0 g/L sucrose and 2.5 g/L gellan gum, and developed into whole plants.

• Journal of Plant Biology
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• v.37 no.3
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• pp.333-341
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• 1994

Effective plant regeneration from immature cotyledons of soybean [Glycine max (L.) Merr.] cultivars was achieved via somatic embryogenesis. Somatic embryogenesis was performed with the cotyledons of immature embryos 14-20 d after flowering. Immature cotyledons of cv. Whangkeum were placed abaxial or adaxial side down on modified MS medium containing 20mg/L 2,4-D. The greatest number of somatic embryos, 1.2 per cotyledon, was produced from those of 4.0-4.9 mm in length which had been placed abaxial side down. Among cvs. Pecking, Whangkeum and Baekwoon, Pecking had the highest embryo induction efficiency with 4.3 somatic embryos per cotyledon in 20mg/L 2,4-D treatment and with 1.0 embryo per cotyledon in 8mg/L NAA treatment. Germinable globular somatic embryos were induced with the highest efficiency, 27.6%, in 20mg/L 2,4-D and were proliferated efficiently on liquid medium containing 10mg/L 2,4-D. The globular somatic embryos developed into germinable mature somatic embryos on medium containing 10 $\mu$M CoCl2, 9% sucrose, and 0.5% activated charcoal. These mature somatic embryos germinated on hormone-free mediu. After transfer to the soil, regenerated plants with seeds were obtained.

• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.39-43
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• 2004

A successful, efficient system for multiple shoot induction and plant regeneration of soybean (Glycine max) was established. Four soybean genotypes were compared for organogenic responses on various media cultured under light conditions. The adventitious shoots (98%, 2.6 shoots/cotyledon) directly from one-day-old cotyledon after germination induced by the hormone treatment and its efficiency was higher than any other conditions. The optimal medium for the induction of multiple shoots from cotyledon in Pungsannamulkong(shoot formation rate, 98%), Lx 16 (83%) and IIpumgeomjeongkong(63%) was MS medium supplemented with 2 mg/L BAP, but for Alchankong(75%), MS medium supplemented with 1mg/L zeatin and 1mg/L IAA, 3% sucrose, 4% Phytagel. Higher root induction (88%) was observed from the shoots placed on rooting medium (hormone-free MS basal). Plantlets were transferred onto the same medium supplemented with 1% activated charcoal for further development. With this treatment, regenerated plantlets were obtained within 7-8 weeks (shoot induction for 4 weeks, rooting and shoot elongation for 3-4 weeks).

• Korean Journal of Medicinal Crop Science
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• v.12 no.5
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• pp.391-396
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• 2004

Optimal culture conditions for efficient in vitro propagation and polysaccharide production of Orostachys japonicus were established. O. japonicus was cultured in media containing various growth regulators and carbon sources. The highest regeneration rate was achieved in 1.0 and $3.0\;mg\;l\;^{-1}$ of 2,4-D concentration, while the lowest was obtained in $10.0\;mg\;l\;^{-1}$ 2,4-D concentration. When different carbone sources were added in the culture medium, plant growth was high in 3% sucrose treatment. The micropropagated shoots were successfully acclimatized in artificial soils and produced comparable amont of polysaccharide compred to parent cultivated plants.

• Horticultural Science & Technology
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• v.32 no.5
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• pp.694-701
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• 2014

Allium hookeri L. (Alliaceae family) is an important ethnomedicinal plant native to the Himalayan region of Asia. The aim of this research was to establish a high-frequency plant regeneration system for in vitro propagation of A. hookeri. Among the tissue types examined, receptacle explants derived from immature flower buds showed the highest regeneration rate of shoots ($93.33{\pm}4.63%$), roots ($76.67{\pm}7.85%$), and calli ($80.00{\pm}7.43%$) when cultured on Gamborg B5 (B5) medium containing $10{\mu}M$ 6-benzylaminopurine (BA) + $1{\mu}M$ naphthalene acetic acid (NAA), $0.5{\mu}M$ BA + $5{\mu}M$ NAA, and $1{\mu}M$ BA + $10{\mu}M$ NAA, respectively. Shoot multiplication was superior when cultured in liquid rather than on solid medium and relatively high concentrations of BA, ranging from 5 to $10{\mu}M$. Efficient bulblet formation following root induction from shoot clumps was achieved with culture in liquid B5 medium containing 7% (w/v) sucrose. Regenerated bulblets were successfully acclimatized to ex vitro conditions with a greater than 95% survival rate. By this method, a maximum of 62 plantlets per receptacle could be propagated within 9 weeks of initial culture. The in vitro propagation system established in this study will promote A. hookeri biotechnology, including large-scale production of healthy and aseptic clones, preserving parental genotypes with desirable traits, and genetic manipulation to enhance medicinal value.

• Journal of Ginseng Research
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• v.36 no.4
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• pp.442-448
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• 2012

Somatic embryogenesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology such as medicinally important plants. Single embryos develop into normal plantlets with shoots and roots. Therefore, direct single embryogenesis derived from single cells is highly important for normal plant regeneration. Here we demonstrate that the cyclic secondary somatic embryogenesis in Panax ginseng Meyer is a permanent source of embryogenic material that can be used for genetic manipulations. Secondary somatic embryos were originated directly from the primary somatic embryos on hormone-free Murashige and Skoog medium, and proliferated further in a cyclic manner. EM medium (one third of modified MS medium [MS medium containing half amount of NH4NO3 and KNO3] with 2% to 3% sucrose) favored further development of proliferated secondary somatic embryos into plantlets with root system. The plantlets developed into plants with well-developed taproots in half-strength Schenk and Hildebrandt basal medium supplemented with 0.5% activated charcoal.

• Journal of Plant Biotechnology
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• v.32 no.2
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• pp.123-128
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• 2005

A series of experiments were performed to establish regeneration system through friable embryogenic callus (FFC) of Lilium longiflorum 'Nellie White'. Only hard and regular callus was induced from bulb scales on medium containing 2.0 mg/L dicamba and $30{\sim}90$ g/L sucrose. The induced hard callus was subcultured on medium with 2.0 mg/L dicamba and 30 g/L sucrose, and used as a material for induction of FEC. In order to induce FEC, induced hard and regular callus was chopped into $1{\sim}2\;mm$ segments, and re-cultured on medium with 2.0 mg/L dicamba and 90 g/L sucrose. FEC was induced from chopped hard calli by the subcultures of two months interval. The induction rate of FEC was enhanced when hard callus was subcultured on same medium. FEC was proliferated more than 5 times on medium with $1.0{\sim}2.0\;mg/L$ dicamba and 90 g/L sucrose. Bulblet differentiation from FEC was very favorable on MS medium supplemented with 0.1 mg/L BA, 1.0 mg/L NAA and 30 g/L maltose, but many differentiated bulblets were changed to vitrificated ones. The differentiation of normal bulblets was most effective on medium containing $0.5{\sim}1.0\%$ activated charcoal and 30 g/L sucrose.

• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.228-235
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• 2010

The establishment of cell suspension culture and plant regeneration of the halophytic Leymus chinensis (Trin.) are described in this study for the first time. Callus induction solid medium containing Murashige and Shoog (MS) basic salt, $2.0\;mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), and $5.0\;mg\;l^{-1}$ L-glutamic acid with $30.0\;g\;l^{-1}$ sucrose and $4.0\;g\;l^{-1}$ gelrite for solidification induced the highest rate of cell division in Type 1 callus among calli of various types. Liquid medium with the same hormone distribution was therefore, used for cell suspension culture from Type 1 callus. Over a 30 d suspension culture at 100 rpm, great amounts of biomass were accumulated, with 71.07% average daily increment and 22.32-fold total fresh weight increment. Comparison of before and after suspension culture, the distribution of different size callus pieces and the maintenance of callus type were basically unaltered, but a slight increase in relative water contents was observed. To induce the potential of plant regeneration, the directly transferring on plant regeneration solid medium containing MS basic salt, $0.2\;mg\;l^{-1}$ $\alpha$-naphthalene acetic acid (NAA), $2.0\;mg\;l^{-1}$ kinetin (Kn), and $2.0\;g\;l^{-1}$ casamino acid and indirectly transferring were simultaneously performed. Even now growth rates of suspension-derived callus on solid medium were approximately half of those of Type 1 callus, but faster somatic embryogenesis was observed. Rooting of all regenerated shoots was successfully performed on half-strength MS medium. All plants appeared phenotypically normal.

• Korean Journal of Plant Resources
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• v.24 no.5
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• pp.591-598
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• 2011

This study was conducted to establish the tissue culture system for Atractylodes plant which is most frequently used in oriental medicine. Root and auxiliary bud of Dachul cv., which is Atractylodes hybrid (A. macrocephala x A. japonica), were used as target tissues for in vitro culture. In root culture, callus induction rate was higher in the treatment of BAP combined with NAA than others, however, 2-iP was more effective for callus proliferation and root induction. Although calli were effectively induced from the root and proliferated in lower concentration of cytokinin combined with higher auxin, root tissue was inappropriate for shoot regeneration. For plant regeneration with axillary bud, BAP combined with NAA was more effective than 2-iP with NAA or IBA. Number of regenerated plant per bud was 3.8, which was highest, and stem diameters was shown as 5.0mm under the conditions of 1 mg/L BAP combined with 1 mg/L NAA. Although, plant height was tend to be higher in 2-iP than BAP, number of the regenerated plant was lower via versus. Furthermore, root proliferation of regenerated plant was more effective in higher concentration of sucrose (7%) than in lower concentration (3%). In results, auxiliary bud was an efficient target tissue for producing regenerated plant of Atractylodes under the conditions of 1 mg/L BAP combined with 1 mg/L NAA and higher concentration of sucrose was effective for root proliferation of regenerated plants.

• Korean Journal of Medicinal Crop Science
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• v.4 no.1
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• pp.78-85
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• 1996

The effects of growth regulators, sucrose and gelling agents were investigated to increase the efficiency of the callus growth and plant regenerarion in tissue culture of Angelica koreana Max. The fresh weight and dry weight of subcultured callus was highest in MS medium supplemented with 1 mg/l 2,4-D. Callus growth was excellent in 2% sucrose, but it was inhibited in propotion to sucrose content. Effect of gelling agents on callus growth was highest on 1.2% agar and 0.4% Gelrite medium, respectively. The browning of callus was protected on the media supplemented with 10 mg/l ABA and 5 or 10 mg/l $AgNO_3$. In the callus induction and growth from the peduncle of immature inflorescence, 2,4-D was more effective than NAA, and the frequency of callus induction was highest as 81.7% in 2 mg/l 2,4-D. Plant was not regenerated from the callus derived from young leaf. Somatic embryos were developed from the surface of callus drived from the peduncle of immature inflorescence in the medium containing 0.5 mg/l 2,4-D, 1 mg/l kinetin, 5 mg/l ABA and 5 mg/l $AgNO_3$. Plants were developed from the matured somatic embryos in the medium supplemented with 0.2 mg/l 2,4-D and 1 mg/l kinetin.