• 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.

• Plant Biotechnology Reports
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• v.2 no.4
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• pp.259-265
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• 2008

Picea koraiensis, called Korean spruce, is an evergreen tree and found mostly in northeast Asia. In this study, plant regeneration via somatic embryogenesis from open-pollinated immature zygotic embryos of nine geno-types of elite trees was established. Immature zygotic embryos were cultured onto RJW medium modified from 505 medium with $21.48{\mu}M$ NAA, $2.22{\mu}M$ BA, and $2.32{\mu}M$ KT. The average frequency for all nine genotypes was 74.2%. Embryogenic calluses of the nine genotypes of elite trees were subcultured on RJW basal medium containing $8.06{\mu}M$ NAA, $1.11{\mu}M$ BA, and $1.16{\mu}M$ kinetin. The calluses of three lines, $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$, were actively proliferated but others were not. Somatic embryogenesis was induced from the embryogenic callus in genotypes of $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$ on RJW medium with ABA and $60g\;l^{-1}$ sucrose. Cotyledonary somatic embryos were subjected to a drying process. The drying of embryos by uncapping the culture bottle for 5 days on a clean bench resulted in a high frequency of germination of somatic embryos (87% in RJW medium). However, plantlet conversion from germinated embryos was greatly reduced and the optimal medium for plant conversion was 1/2 WPM or 1/2 BMI medium. In conclusion, we have, for the first time, established a plant regeneration system via somatic embryogenesis in the Korean spruce, which can be applied for rapid micropropagation of elite trees.

• Journal of Plant Biology
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• v.19 no.2
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• pp.45-48
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• 1976

Anthers of cultivated Paeonia albiflora were cultured on media supplemented with various combinations of growth regulators. Although the number of anthers with emerged calluses were very few, in the sectioned anthers were found many multinucleate, 2-celled, or multicellular microspores, the one-celled multinucleate microspores being most abundant in number, and the multicellular ones the least. In 2-celled or 3-celled microspores two kinds were observed: one is ordinary one with single nucleus in each cell, and the other is multinucleate one. Majority of the 2-celled microspores was found to be of equational-division irrespective of whether they were multinucleate micropores or ordinary nonmultinucleate ones.

• Journal of Plant Biotechnology
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• v.46 no.4
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• pp.297-302
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• 2019

Hibiscus syriacus L., the national flower of Korea, is a commonly grown ornamental species found in parks, gardens and along roadsides in Korea. This study sought to establish a plant regeneration system of H. syriacus 'Nanpa' using mature leaves as an explant. Sterilized mature leaf explants were cultured on modified MS medium with combinations of NAA and BAP at various concentrations for 6 weeks. Among the various media evaluated, modified MS media containing 0.46 mg/L NAA and 0.5 mg/L BAP (CI-1) or 0.92 mg/L NAA and 1 mg/L BAP (CI-4) were the most effective for callus formation. Whitish-yellow calluses were observed on CI-1 medium, while green calluses formed on CI-4 medium. The whitish-yellow callus derived from CI-1 medium showed a higher frequency of shoot induction than the green callus derived from CI-4 on modified MS medium containing 0.5 mg/L BAP. Among the various media evaluated in this study, modified MS medium containing 0.46 mg/L NAA and 0.5 mg/L BAP was the most optimal for shoot-forming callus production. Our findings show that mature leaves of H. syriacus 'Nanpa' can be used for in vitro plant regeneration, and the regeneration system described here may be a powerful tool for molecular breeding of H. syriacus 'Nanpa' in the future.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.9
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• pp.5628-5636
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• 2014

Currently, many countries have an interest in developing cosmetics materials using native plants. In this aspect, there is increasing need to develop cosmetics materials using native plants in our county. In the present study, calluses were induced from Artemisia annua Linne, which was highlighted because of its useful effects, such as anti-cancer, anti-fungal and anti-inflammation. Water and ethanol extractions were performed from the calluses of Artemisia annua Linne. After the mass production of Artemisia annua Linne's calluses, water and ethanol extraction was performed to examine its functional roles in healing wounds and inflammation. The differences in the effective elements were observed in the ethanol extract. The callus showed anti-inflammation activity through the suppression of the inflammation-related gene, COX-2, and ethanol extracts showed their ability to heal wounds. Overall, these results suggest that the extract of Artemisia annua Linne's calluses is a natural and environment-friendly material, and can be used as medical supplies associated with anti-inflammation and healing wounds.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.146-153
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• 2012

In this study, the antioxidant property of leaf and callus extracts of five selected in vitro grown Ocimum species (Ocimum sanctum, Ocimum kilimandscharicum, Ocimum gratissimum, Ocimum basilicum, and Ocimum americanum) and their respective callus extracts was investigated. The callus cultures were successfully initiated on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (1mg L) combined with different concentrations (0.1-0.4 mg L) of kinetin as plant growth regulators. Total phenolic contents were estimated using the Folin-Ciocalteu reagent. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power, $Fe^{2+}$ chelating activity, and ${\beta}$-carotenelinoleic acid bleaching assays were used to determine the biological effects of the extracts. Interestingly, all the callus extracts exhibited significant (p<0.05) increase in phenolic contents and antioxidant activity. Furthermore, a liner correlation was obtained between the total phenolic contents and free radical scavenging activity ($R^2$ = 0.783). The extracts of leaves and calluses of Ocimum species exhibited activity in all the in vitro antioxidant assays, but its extent was less potent that the positive controls butylated hydroxyl anisole (BHA) and ascorbic acid. A higher accumulation of phenolics in the callus extracts suggests that isolation of high-concentration materials with antioxidant activivity is possible from in vitro callus cultures rather than field-grown plant organs. Furthermore, these extracts may be used as an effective preservative in the food industry.

• Korean Journal of Soil Science and Fertilizer
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• v.12 no.1
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• pp.43-46
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• 1979

Curing the studies on the mechanism of the nodule formation and the infection of rhizobia using the continously cultured plant cell tissues, it was found that some calluses possess high nitrogen fixation activity. This experiment was conducted to know the differences among the calluses and the Rhizobia. The results obtained were as follows; 1) In a single inoculation the nitrogenase activity of soybean cell cultures-rhizobium was moderately higher than non-leguminous cell cultures, however, in the mixing inoculation that was reversely found. 2) Host factor, which was characteristics for the nodule formation and the nitrogenase activity, was not appeared generally in the plant cell cultures except for Kuamusume and Toyozuzu in soybean, and Datura in non-leguminous cell cultures. 3) In the 012 rhizoblium cultured on soybean cell cultures and in the 010, 023 and 024 rhizobia cultured on non-leguminous cell cultures the nitrogenase activity higher than the others.

• Journal of Plant Biology
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• v.31 no.4
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• pp.309-315
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• 1988

Leaf mesophy11 protoplast cultures from six Nicotiana species, N. debneyi, N. rustica, N. amplexicaulis, N. glauca, N. glutinosa, and N. sylvestris were carried out. When we reduced the NH4NO3 and Fe.EDTA concentration to 1/3(7 mM) and 1/10(10$\mu$M) from the Murashige and Skoog medium respectively, cell division of the protoplasts was efficiently induced in four Nicotiana species, N. debneyi, N. rustica, N. amplexicaulis and N. glauca. However, other two species, N. glutinosa and N. sylvestris were failed in inducing cell division at the same culture condition. The protoclone calluses derived from four Nicotiana species were consequently regenerated on a MS basal medium supplemented with the appropriate auxin and cytokinin.

• Journal of Plant Biology
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• v.37 no.1
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• pp.27-36
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• 1994

For establishing a transformation system of rice, an efficient introduction of foreign genes into embryogenic cell suspension by particle bombardment was conducted. The particle inflow gun based on the acceleration of DNA-coated tungsten particles using pressurized helium was constructed for delivery of DNA into rice cells. Several bombardment parameters were optimized using the transient expression of GUS gene. The conditions that gave the highest GUS gene expression of about 1000 blue spots per g fresh weight of bombarded cells include treatment of the cells with 0.5 M osmotic pressure, and use of the 410 kPa helium, 110 mm target distance, 13 mm syringe filter holder and 5 $\mu$L DNA/tungsten mixtures. It was also confirmed that rice actin promoter-intron construct gave the highest expression of all promoter-sequences studied. Eight weeks after the bombardment, stably transformed calluses were obtained on the selection medium containing 100 mg/L G418 and showed the strong activity in in situ GUS assay.

• Journal of Plant Biology
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• v.29 no.3
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• pp.197-205
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• 1986

Leaf mesophyll protoplasts from five cultivars of tobacco (N. tabacum L.) were cultured. The protoplasts did not survive in culture medium containing Murashige and Skoog inorganic salts for over 6 days. NH4NO3 and FeSO4.Na2EDTA concentration of Murashige and Skoog medium were toxic in tobacco leaf mesophyll protoplast culture. Therefore we investigated optimum condition in Murashige and Skoog medium. High plating efficiency was obtained by reducing the concentrations of NH4NO3 and FeSO4.Na2EDTA to 1/3 and 1/10, respectively, on the supplemented with 5$\mu$M IAA, 0.5 $\mu$M 2.4-D 5 $\mu$M BAP. Plants were regenerated from protoplast-derived calluses.