• Korean Journal of Medicinal Crop Science
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• v.13 no.1
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• pp.57-62
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• 2005

Salt-tolerant transgenic Panax ginseng plants were produced by introducing the SAL1 geue (3'(2'), 5'-bis-phosphate nucleotidase) that confers tolerance to the salts through Agrobacterium tumefaciens co-cultivation. Cotyledon explants of immature ginseng zygotic embryos cultured on Murashige and Skoog medium lacking growth regulators formed somatic embryos directly with below 10%, but the 74% tranformation rate were observed at the treatment of phytohormone with 1.0 mg/l 2,4-D and 0.5 mg/l kinetin. Somatic embryos were initially cultured on MS medium supplemented with 250 mg/l cefotaxime for 3 weeks and subsequently subcultured five times to a medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime. Upon development into the cotyledonary stage, these somatic embryos were transferred to on the medium containing 50 mg/l kanamycin and 10 mg/l gibberellic acid to induce germination and strong selection. Integration of the transgene into the plants was confirmed by polymerase chain reaction with specific primers. The ginseng transformants with well-developed shoots and roots were successfully acclimatized in a greenhouse when they were planted in soil.

• Applied Biological Chemistry
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• v.27 no.2
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• pp.129-134
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• 1984

Change of protein component in soybean sprout grown at four temperatures was investigated by polyacrylamide gel electrophoresis. Main bands were identified using purified seed globulins. Electrophoretogram showed 5 main bands (a. b, c, d, and p) and 10 minor bands in seed and maximum number (19) of bands (8 main band including 0 and 11 minor) at 4th day after germination in cotyledon. All bands appeared in axis protein but resolution was poor. In cotyledon, a component (most rapidly) and b+c+d component decreased while o+p component and other minor components were increased at 6th day and decreased thereafter. In axis all components increased rapidly, especially in minor components and b+c+d component. High growing temperature accelerated decrease in cotyledon and increase in axis of protein, especially for 11S. The a component was identified as 7S, b+c+d as 11S and o+p as 2S globulin.

• Journal of Korean Society of Forest Science
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• v.97 no.1
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• pp.127-133
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• 2008

This study was conducted to establish the optimal condition for plant regeneration and acclimatization from somatic embryos of Panax ginseng. Cotyledon segments of Panax ginseng produced primary and secondary somatic embryos when cultured on MS and WPM media supplemented with 7% sucrose. To induce plantlet conversion, cotyledonary somatic embryos were cultured on WPM solid medium with $GA_3$ at various concentrations (1~30 mg/L) for 4 weeks. Plantlets were transferred to 1/2 WPM solid medium with $GA_3$ at various concentrations (0~5 mg/L) and 0.5% activated charcoal for shoot and root elongations. Elongated plantlets further developed into well-developed leaf and root system on 1/3 SH medium with 0.5% activated charcoal under ventilation condition for 5 months. The highest survival rate to soil was 75% when plantlets were regenerated on 1/3 SH medium without sucrose under ventilation condition.

• Journal of Ginseng Research
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• v.27 no.1
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• pp.37-42
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• 2003

For study about introduce of gene connected with disease and transformation system of gingseng, chitinase gene cloned from soybene and disease resistant gene were carried out for expression and transformation of plant using Agrobacterium. The disease resistance gene(DR-49), 35S-35S-AMV, has been constructed. The disease resistance gene and chitinase gene were introduced into the binary vector pRD 400, which were mobilized into Agrobacterium tumefaciens faciens strain MP 90 and LBA 4404 harboring disarmed Ti-plasmid. As a result of induce transformants using ginseng embryo and petiole, multi shoots were formed on MS medium supplemented 1 mg/ι 2,4-D and 0.5 mg/ι kinetin. Also transformation by cotyledonwas effective on MS medium supplemented 1 mg/ι 2,4-D and 0.5 mg/ι kinetin, transformation percent of disease resistant gene and chitinase gene were showed 18%, 14% respectively. As transformed tissue is under pre-embryoid condition, normal shoot is required through the process of matured embryo.

• Journal of Ginseng Research
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• v.37 no.3
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• pp.332-340
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• 2013

In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.31 no.2
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• pp.123-128
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• 1986

The characteristics such as characters of seed, fruit and endosperm, development of embryo, dehisced and germination rate of seeds which harvested at 10 days intervals from 20 days to 60 days after pollination were determined to clarify the optimum time for seed production of ginseng. The sizes of length, width and thickness of fruits, seeds and endosperms investigated on 20 days after pollination were not largely different from those that harvested after 60 days. But the weights of fruits and seeds were increased along with the delay of harvesting time. The seeds which harvested from 30 to 60 days showed comparatively high dehisced and germinated rates, especially 40 days in dehisced rates and 50 days in germinated rates. The embryos were rapidly grown in early stage. Lengths of the embryo on 20 days after pollination were approximately grown to 30 ${\mu}{\textrm}{m}$. And after 30 days pollinated, embryos were not only more rapidly grown to 213 ${\mu}{\textrm}{m}$ but also cotyledon premodia were developed in this time and completely formed after 40 days.

• Korean Journal of Medicinal Crop Science
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• v.8 no.1
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• pp.14-20
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• 2000

This experiment was conducted to investigate the effects of length of storage period under low temperature, $CO_2$ enrichment and addition of plant growth regulators in Murashige and Skoog medium on the plant regeneration of Korean ginseng (Panax ginseng C. A. Meyer). Seeds were treated for 60 and 80 days respectively under $5^{\circ}C$ environment. 2500ppm of $CO_2$ was enriched by ventilation. Three plant growth regulators added to the medium were Indolbutyric acid, Benzyladenin and Gibberellic acid (GA3). The result indicated that : The capacity of differentiation was higher in the aged cotyledons from the seeds treated for 80 days under low temperature condition than in those treated for 60 days. $CO_2$ enrichment had stimulating effects on the growth and development of shoot primordium significantly but less effects on the formation of adventitious buds. From one zygotic embryo hundreds of plantlets were differentiated. $CO_2$ enrichment had effects on the formation of both indirect somatic embryo and direct somatic embryo. Indirect somatic embryo showed little growth and differentiation, being undifferentiated vascular stele and epicotyl. Direct somatic embryos were formed on the epidermis of backside basal part of cotyledon. Those embryos developed to whole plant having latent bud.

• Applied Microscopy
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• v.9 no.1
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• pp.57-69
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• 1979

The ultrastructural changes of embryo and endosperm cells were observed during the green fruit with embryo about $250{\mu}$ long to germination. 1. In the embryo cells of green fruit with embryo about $250{\mu}$ long, mitochondrial cristae and plastid are undifferentiated and dictyosome are occasionally observed. There are electron-opaque globoids in the vacuole and a lot of spherosomes in the outer layer of smooth endoplasmic reticulum. Endosperm is filled with spherosomes and electron-opaque protein bodies surrounded by spherosomes, and due to these, other organelle are not observed. 2. In the embryo cells of seeds with red seed coat, mitochondrial cristae are well developed, electron-opaque globoids increased, and vacuoles are enlarged. In the endosperm, however, spherosomes increased, protein bodies are enlarged, and electron-opaque globoidal crystals are dispersed within them. 3. In the procambium and epicotyl cells of dehiscent seed, Golgi vacuoles and vesicles are well developed, and mitochondrial cristae are also well differentiated. Spherosomes are numerously present and radicle cells, peripheral cells of hypocotyl, and vacuoles of cotyledon are well differentiated. Endosperm is filled with spherosomes containing electron-opaque granules and protein bodies are surrounded by a single membrane. There are acid phosphatase around globoids and spherosomes. 4. At the time of seeding, spherosomes markedly increased in the outer layer of cotyledon and protein bodies are also observed. Cell organelles are differentiated and plastids containing starch are also present. 5. In the outer $2{\sim}3$ layers of cotyledons, radicle cells, and peripheral cells of hypocotyl during post-seeding to germination, spherosomes and plastids with starch increased, and mitochondria and microbodies are also found around the nucleus of embryo cells. With approaching, the germination stage, in the endosperm contacting with embryo, vacuoles are well differentiated but spherosomes decreased. There increased electron-opaque materials within vacuoles. In other endosperm, with the decrease of spherosome, mitochondria increased and electro n-opaque globular bodies are formed and gradually increased. The outer layer of protein bodies are reduced while electron-transparent portions are enlarged and fused together to occupy the outer layer where small particles are formed. 6. In the endosperm of germination stage, spherosomes decreased while protein bodies, are fused together to form 2 or 3 within a cell.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.111-115
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• 2002

Taraxacum platycarpum has been used as a medicinal plant. We investigated optimal condition for efficient plant regeneration through adventitious shoot formation on medium with various kinds of growth regulators. Adventitious shoot formation was achieved when cytokinin was used alone. Shoot formation was higher on MS medium containing 2 mg/L BAP compared to that with 2 mg/L kinetin and 2 mg/L 2-ip. Among root, hypocotyl and cotyledon, roots were the best explant for the adventitious shoot induction. Adventitious shoot formation from roots declined markedly by the combination of both 0.1 mg/L NAA and 2 mg/L BAP, while shoot formation from cotyledons was stimulated by the same combination. Root formation from the regenerated shoots was achieved on 1/3MS medium containing 0.2 mg/L NAA. Regenerated plantlets was acclimatized and transplanted to the soil, showing 100% survival.