• Korean Journal of Medicinal Crop Science
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• v.10 no.2
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• pp.116-119
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• 2002

A protocol has been developed for differentiation of adventitious shoots directly from leaf segments of Tetragonia tetragonoides O. Kuntze. Murashige and Skoog (MS) medium supplemented with 2 mg/L $N^6-benzyladenine$ (BA) and 0.5 mg/L ${\alpha}-naphthaleneacetic$ acid (NAA) supported the induction of adventitious shoots from leaf explants. Adventitious shoots were multiplied by subculturing on the double strength MS (2MS) medium supplemented with 0.5 mg/L NAA and 2 mg/L BA. Shoots were rooted on MS basal medium without any growth regulators.

• Journal of The Korean Society of Grassland and Forage Science
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• v.18 no.1
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• pp.69-76
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• 1998

This study was conducted to construct of the transgenic plants wliich are resistant to oxidative stresses including ozone with B. mpestris cytosolic glutathione reductase cDNA using the binary vector system of Agrobacterium tumefaciens. The 1.8kb B. campestris cytosolic GR cDNA was subcloned into the unique Sma I site of the plant transformation vector pBKSI- I, downstream of the constitutive CaMV 35s promoter and upstream of the nos termination sequence, in place of the uidA (GUS) reporter gene. The resulting plant transformation vector, pBKS-GRI, was introduced into A. tumefaciens LBA4404 by two cycles of tkeze-thaw method. The B. nqus cotyledonary petioles were transformed by the Agrubaferium harboring pBKS-GRI. Transformed shoots were induced and selected on regeneration medium supplemented with kanarnycin. The shoot formation was increased remarkably by addition of Ag$NO_3$, in MS media. The transgenic plants were analyzed for the presence of the B. campestris GR gene by Southern blot analysis and it was confirmed that a foregin gene was stably integrated into the genomes of B. nqus plants.

• Horticultural Science & Technology
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• v.29 no.3
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• pp.260-266
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• 2011

An efficient protocol was established for high frequency somatic embryogenesis through a callus culture of Coelogyne cristata. The best frequency of callusing was obtained from a PLB segment (3-5 mm) cultured on MS medium supplemented with coconut water (CW) and a combination of both 3 $mg{\cdot}L^{-1}$ of 2,4-D and BA. When the calli were sub-cultured on the MS medium without any PGRs, the average number of somatic embryos were higher than those with PGRs treatment. NAA is the most critical factor among PGRs, which dramatically hindered for the formation of a somatic embryo. The efficacy of the addition of coconut powder (CP) for somatic embryogenesis was almost the same in all treatments. However, the number of somatic embryos formed distinctly depended on age of the callus. The somatic embryos converted into healthy plants with well-developed shoots on the same medium. Plantlets showed the best responses of root and shoot growth when transferred to $\frac{1}{2}$ MS medium containing 1.5 $g{\cdot}L^{-1}$ of activated charcoal. All plants with above 3.0-cm-high were successfully acclimatized in the greenhouse.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.285-289
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• 1997

Leaf and stem explants of Sedum erythrostichum Miq. were cultured on MS medium supplemented with various combinations of growth regulators. After two weeks of culture, 100% of the leaf explants formed calli on medium containing 2.0 mg/L 2, 4-D and 1.0 mg/L BA. Callus proliferated when subcultured on medium containing 2.0 mg/L 2, 4-D and 1.0 mg/L BA. Numerous adventitious buds were regenerated from callus cultured on the medium containing 2.0 mg/L NAA and 1.0 mg/L BA. Root formation from shoot was occurred on the MS basal medium containing 1.0 mg/L IAA.

• Journal of The Korean Society of Grassland and Forage Science
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• v.23 no.2
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• pp.95-100
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• 2003

The alfalfa (Medicago sativa L.) callus was induced from seeds on SH medium contained $3\;mg/{\ell}$ of 2,4-D. Several regenerated alfalfa plants and many shoots were obtained by procedure of Kim et al. (1999); 1) incubation for 28~30 days on SH medium added $5\;mg/{\ell}$ of NAA and $2\;mg/{\ell}$ of Kinetin, 2) incubation for 3~5 days on SH medium added $11\;mg/{\ell}$ of 2,4-D and $1\;mg/{\ell}$ of kinetin. 3) incubation for 21~25 days on SH medium added $1.6\;g/{\ell}$ of ammonium sulfate and $5.75\;g/{\ell}$ of proline. To increase of root induction ratio on plant regeneration process of alfalfa, root induction ratio was examined on 8 kinds of medium, containing different amount of hormone and SH salt. Root induction ratio was higher on SH medium contained IBA than SH basal medium. In case 1.5 mg of IBA was added in SH medium, root induction ratio was the highest to 56.0% in this study. On the other hand, root induction ratio was higher on SH medium diminished SH salt amount to half volume and addition of IBA makes high root induction ratio, too. Thus, we conclude that the medium for root induction of alfalfa may be added $1.5\;mg/{\ell}$ of IBA and diminished SH salt amount to half volume.

• Korean Journal of Plant Tissue Culture
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• v.27 no.2
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• pp.101-107
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• 2000

The explants of yacon (Polymnia sonchifolia Poeppig & Endlicher) were cultured to invest th8e dedifferentiation condition, and formative callus from leaf was cultured to find the regeneration and micro-crown bud formation. Basal MS medium was more effective to form callus than 1/2 MS and B$_{5}$ medium. Calli formations from leaf, petiole and lateral bud were more effective on MS medium supplemented with 1.0, 2.0 mg/L 2,4-D and 0.2, 0.4 mg/L kinetin or BA than 1.0, 2.0 mg/L NAA and 0.2, 0.4 mg/L kinetin or BA. Formative callus from leaf was proliferated about 70% on medium supplemented with 1.0 mg/L BA. When callus was proliferated, 63% regeneration rate was shown on medium supplemented with 1.0, 2.0 mg/L BA in case of subculture for 3~4 months but was not shown on medium supplemented with 1.0, 2.0 mg/L kinetin. Micro-crown bud formed as addition of BA at 3~4 months after callus culture and then was obtained many at 5~6 months, it was most formed about 82% on medium supplemented with 5 mg/L BA. Rate of micro-crown bud formation was increased as more over 5 mg/L BA concentration, when this time, however, shoot had thick leaves and short internodes, and then withered before long, Micro-crown bud was formed about 88.0% on medium supplemented with 5% sucrose, that was more increased 28% than with 3% sucrose. The buds of crown bud between harvested in field and formed in vitro were difference only in size, but both were similar in shape according to histological view.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.69-76
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• 2011

This study describes the effect of myo-inositol on sustained cell division and plant regeneration from cotyledon-derived protoplast of cabbage (Brassica oleracea var. capitata). Freshly isolated protoplasts were cultured in modified Murashige and Skoog (MS) medium removed ammonia ions and containing $0.4\;mg\;l^{-1}$ thiamine HCl, $100\;mg\;l^{-1}$ myo-inositol, $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and several concentrations of myo-inositol (2, 4, 6, 8, 10% (w/v)) as an osmotic stabilizer. After 3 weeks of culture in the dark at $25^{\circ}C$, the plating efficiency of cabbage protoplasts reached to $22.5{\pm}2.9%$ when cultured in modified MS medium supplemented with $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and 8% (w/v) of myo-inositol at a density of $2{\times}10^5$ protoplasts/ml. Rapidly growing cell colonies after 3 weeks of culture were transferred to the same culture medium removed osmoticum. To induce shoot regeneration from calluses, calluses with about 2 mm in diameter were transferred to the MS medium containing $2\;mgl^{-1}$ BA and $0.5\;mgl^{-1}$ NAA. After further three weeks of incubation onto the medium in the light, green shoots were formed on the surface of calluses at a frequency of 30%. Upon transfer to half-strength MS basal medium, roots were formed onto the bottom of regenerated shoots without auxin treatments. These regenerated plantlets were successfully acclimatized to soil transfer, grown to normal mature plants. The cabbage protoplast culture system established in this study could be applied for production of somatic hybrids or cybrids by asymmetric protoplast fusion and mass proliferation of elite somatic clones of cabbage.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.197-203
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• 1999

The albino plants of tobacco (Nicotiana tobacum L. cv. BY-4) were isolated from seed populations that were induced by heavy-ion ($^{14}N$) beam irradiation to proembryo and the in vitro growth and differentiation have been characterized. The in vitro cultured albino plants showed significant reduction of chlorophyll content and possessed larger number of stomata on both upper and lower epidermis than that of wild-type plants. Stem growth of the mutants remained dwarfed, however, the internode recovered its normal length after GA$_3$ treatment (10.0mg/L) on the MS medium containing sucrose under continuous light. When explants of leaf blades of albino plants were cultured, multiple shoots formed directly on MS medium containing 1.0mg/L of BAP or kinetin and a large number of calli were induced on the MS medium containing 1.0mg/L NAA or 1.0 mg/L 2,4-D. The albino calli regenerated multiple albino plantlets in the MS medium containing 0.1mg/L NAA + 1.0 mg/L BAP. No significant differences between the wild-type and albino plants were detected in the multiple shoot induction, callus formation from the explants and the plantlets regeneration from calli. In addition, albino plants have a similar organogenesis Pattern to that of the wild-type in the media with different combinations of NAA (0 to 5.0mg/L) and BAP (0 to 5.0mg/L) treatment. These results indicate that the albino mutant has the same normal regeneration ability as that of wild-type, although the mutant has lost functions in photosynthesis, such as pigmentation.

• Korean Journal of Plant Resources
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• v.19 no.5
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• pp.634-639
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• 2006

In order to develop a disease-resistant root stock for the growth of watermelon, an efficient regeneration system of the gourd(Lagenaria leucantha Duch.) inbred line GO701-2 via organogenesis was established in this experiment. Using proximal parts of cotyledon explant excised from germinated seedling in vitro, maximum adventitious shoot formation (39%) was achieved on MS medium where cytokinin (BA) and auxin (IAA) were added at a concentration of 3mg/L and 0.1mg/L, respectively. Roots of the elongated shoots were successfully formed on MS medium without adding any plant growth regulators. The cucumber CsGolS1 gene known as a resistance gene against biotic and abiotic stresses, was constructed into the binary vector pBI121 under the control of CaMV 35S promoter. When the gene was introduced into the genome of gourd by Agrobacterium-mediated transformation, putative transgenic plants were obtained with the transformation efficiency of approximately 20 percent.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.184-189
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• 2000

Callus was induced from the petioles of scented-geranium (Pelargonium graveolense) in MS medium containing various concentrations of plant growth regulators. The highest frequency of more than 70% of callus was induced in 2 mg/l NAA and 0.5 mg/l BAP or 2 mg/l 2,4-D and 0.5 mg/l BAP combined treatment, while not in 2,4-D, NAA or BAP alone. When the callus was transferred to the MS medium containing 0.05 mg/l NAA and 0.5 mg/l BAP, were highest intensity of shoot formation, 14 shoots/callus, was induced after 5 weeks. The highest rooting was observed on hormone-free rooting media from the regenerated shoots after 3 weeks, indicating that the regeneration from geranium callus might be possible by changing the hormone ratios. Peroxidase (POD) specific activities in callus induced from 2 mg/l NAA and 0.5 mg/l BAP were higher than those of 2 mg/l 2,4-D and 0.5 mg/l BAP callus during the entire culture periods. POD isozyme C3 was the main cathodic POD isozyme expressed in NAA and BAP callus, while C1 was the main in 2,4-D and BAP callus. However, anodic POD isozymes, A1, A2 and A3 were expressed with similar activities in both hormone combinations.