• Journal of Plant Biotechnology
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• v.1 no.2
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• pp.101-107
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• 1999

The leaf blade-segments of albino tobacco (Nicotiana tabacum L.) were cultured on MS media containing different concentrations of BAP (0, 0.4, 2.2, 4.4, 22.2 ${\mu}{\textrm}{m}$) with or without NAA (0, 0.5, 2.7 ${\mu}{\textrm}{m}$). Multiple shoots were induced on the media containing 0.4 to 2.2 ${\mu}{\textrm}{m}$ BAP. The best condition for multiple shoot induction with root formation was MS media containing 4.4 ${\mu}{\textrm}{m}$ BAP and 0.5 ${\mu}{\textrm}{m}$ NAA. The regenerated albino plants showed a significant reduction in accumulation of chlorophylls and carotenoids. The drastic reduction of the pigments content was associated with the distinct alterations in gene expression in the albino plants. firstly, the expression of plastid genes, such as rbcL, psbA, 165 rDNA and 235 rDNA, was reduced at the level of transcripts in the regenerated albino plants. Secondly, the alteration of structure of the plastid genes was not detected in the albino plants. However, the copy number of the plastid genes whose transcription level was reduced greatly was increased approximately two-fold, although the transcriptions of nuclear gene (255 rDNA) showed the wild-type level.

• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.84-88
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• 2010

To establish the system of In vitro plant regeneration, the floral bud and leaf explants of Sedum rotundifolium were cultured on the MS media supplemented with different concentration of 2,4-D, NAA, and BA. The callus induction was more effective in the floral explants than the leaf explants, and was the best on MS medium containing 1.0 or 2.0 mg/L 2,4-D and 1.0 mg/L BA. The highest numbers of shoots were regenerated when callus were cultured on MS medium containing 2.0 mg/L 2,4-D and 1.0 mg/L BA for 8 weeks. The normal root formation from shoot was effective on the MS medium containing IAA alone. The regenerated plantlets were transferred to the pot and acclimatized successfully.

• Proceedings of the Botanical Society of Korea Conference
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• 1987.07a
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• pp.213-237
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• 1987

In vitro flowering system may minimize the confounded influence of non-floral meristem parts of plants in studying the relationship of a given treatment and flowering responses. We have induced flower buds from plantlets regenerated from zygotic embryo-derived somatic embryos of ginseng, which circumvented the normal 2-year juvenile period before flowering. The result suggests that the adulthood of ginseng root explants in the experiment previously conducted by Chang and Hsing (1980; Nature 284: 341-342) is not prerequired to flowering of plantlets regenerated through somatic embryogenesis. We have also induced flower buds from elongated axillary brandches from cotyledonary nodes by culturing ginseng zygotic embryos, seedlings, and excised cotyledonary nodes. It was found that 6-benzyladenine (BA) supplemented to the medium was essential for flowering, whereas abscisic acid (ABA) was inhibitory. Gibberellic acid(GA3) was also required for flowering when ABA was present with BA in the medium. The results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respective-ly, in the induction of flowering of ginseng. Tran Thanh Van (1980; Int. Rev. Cytol., Suppl. IIA: 175-194) has developed the "thin cell layer system" in which the induction of shoots, roots, or flower buds from epidermal layer explants were controlled by culture conditions and exogenous growth regulators in the medium, Utilizing the thin cell layer system, Meeks-Wagner et al. (1989; The Plant Cell 1: 25-35) have cloned genes specifically expressed during floral evocation. However, the system is too tedious for obtaining a sufficient amount of plant materials for biochmical and molecular biological studies of flowering. We have developed a garlic callus culture system and one obvious advantaging over the thin cell layer system is that an abundant cells committed to develope into flower buds proliferate. When the above cells were compared by two-dimensional gel electrophoresis with those which have just lost the competence for developing into flower buds, a few putative proteins specific to floral evocation were detected. The garlic callus culture system can be further explored for elucidation of the molecular biological mechanism of floral evocation and morphogenesis.hogenesis.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.10b
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• pp.21-21
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• 2003

An efficient plant regeneration system was developed for Hordeum vulgare L. cv Baegdong - an important Korean cultivar. The protocol was based on a series of experiments involving the sizes of mature embryos and the culture media. The embryo size is found to be critical for the establishment of embryogenic callus. Embryos of 1.1-1.5 mm size showed a much higher ability to produce embryogenic callus capable of regenerating green plants. The auxins picloram and dicamba proved effective in inducing callus from mature embryos. 2.5 mg $I^{-1}$ dicamba and 4.0 mg $I^{-1}$ picloram in Murashige and Skoog's (MS) medium was optimum for the induction of primary callus. The induced primary callus was loose and friable which ultimately developed into creamy white and compact callus after transferring into the fresh medium. Multiple shoots were induced in the MS medium supplemented with 6.0 g $I^{-1}$ maltose, 20 mg $I^{-1}$ sorbitol, 0.5 mg $I^{-1}$ 2,4-D and 1.0 mg $I^{-1}$ kinetin and the rate was 6.5 shoots per embryo. Regenerated plants were hardy and developed roots rapidly in the medium containing 0.2 $I^{-1}$ IBA. This efficient plant regeneration system provides a foundation for generating transgenic plants of this important barley cultivar.

• Journal of Plant Biotechnology
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• v.6 no.4
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• pp.241-246
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• 2004

The bulb scales and shoot sections ($7\;\cal{mm}\;\times\;15\;\cal{mm}$) of Lilium oriental hybrid 'Casablanca' were cultured to compare bulblet growth in vitro. Shoots were induced from in vitro grown bulbscales on MS medium with $1.0\;\cal{mg/L}\;BA,\;0.5\;\cal{mg/L}$ IAA, and 30 g/L sucrose. The regenerated shoots were cut into shoot sections, and cultured on MS medium with $2.0\;\cal{mg/L}\;BA,\;0.5\;\cal{mg/L}$ IAA and 30 g/L sucrose for shoot proliferation. Culture of shoot sections stimulated bulblet growth significantly than the bulb scales on MS medium with 60 g/L sucrose. However, the bulblets from shoot sections did not reach ideal size to produce stems with several leaves. Therefore, liquid medium was added into the same vessels to stimulate bulblet growth further. After shoot sections were cultured on MS medium with 60 g/L sucrose and 2 g/L activated charcoal for two months in dark, $20\;\cal{ml}$ liquid media containing various concentrations of sucrose and MS salts were added. Two months later, the added liquid medium stimulated bulblet growth remarkably as compared to bulblets grown without added liquid medium. The added $25\;\cal{ml}$ liquid medium containing 120 g/L sucrose and double strength of MS salts were the most effective for growth of in vitro bulblets. More than $94\%$ bulblets produced by this method sprouted stems with several leaves after cold treatment at $5^{\circ}C$ for three months.

• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.1-6
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• 1999

In order to obtain plantlet derived by anthers, the anthers of Lilium asiatic hybrid 'Gran Paradiso' were cultured on Murashige and Skoog's medium supplemented with various combinations of auxin and cytokinin. The most suitable pollen stage of anther culture for the callus induction was 3 days before anthesis at the early to late binucleate stage. Organogenic calli were induced on MS medium supplemented with 5.0 mg/L 2,4-D alone and the combination of 1.0 mg/L 2,4-D and 1.0 mg/L kinetin, however, the combination of NAA and BA was more effective than that of 2,4-D and kinetin on plant regeneration through organogenesis. Shoots were formed from the induced callus on the medium with 0.5 mg/L NAA and 1.0 mg/L BA after 180 days of culture. Multiple shoots with 3-4 leaves, roots, and bulblets were formed on the medium with the combination of 2.0 mg/L NAA and 2.0 mg/L BA after 250 days of culture. The chromosome from root tip of the regenerated plantlet showed the diploid (2n=2x=24). Diploid plants were transferred to the pots and all plants were flowered in two years.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.77-85
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• 2004

Development of efficient plant regeneration and genetic transformation protocols (using the Particle Inflow micro-projectile Gun and the shoot-tips as target tissue) of Sorghum bicolor (L.) Moench in terms of expression of the reporter gene, $\beta$-glucuronidase(uidA) is reported here. Two Indian cultivars of sorghum were used in the study, viz. M-35-1 and CSV-15. Plant regeneration was achieved from one-week-old seedling shoot-tip explants via multiple-shoot-clumps and also somatic embryos. The multiple-shoot-clumps were produced on MS medium containing BA (0.5, 1.0 or 2.0 mg/$L^{-1}$), with biweekly subculture. Somatic embryos were directly produced on the enlarged dome shaped expansive structures that developed from shoot-tip explants (without any callus formation) when cultured on MS medium supplemented both with BA (0.5, 1.0 or 2.0 mg/$L^{-1}$) and 2,4-D (0.5 mg/$L^{-1}$). Whereas each multiple-shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing 1.0mg/$L^{-1}$ IBA and successfully transplanted to the glasshouse and grown to maturity with a survival rate of 92%. The plant regeneration efficiency of both the genotypes were similar. After the micro-projectile bombardment, expression of uidA gene was determined by scoring blue transformed cell sectors in the bombarded tissue by an in situ enzyme assay. The optimal conditions comprising a helium pressure of 2200 K Pa, the target distance of 11 cm with helium inlet fully opened and the use of osmoticum have been defined to aid our future strategies of genetic engineering in sorghum with genes for tolerance to biotic and abiotic stresses.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.335-342
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• 2017

Purple passion fruit (Passiflora edulis Sims) is one of the introduced tropical plants, an increasing interest has arisen due to its distinctive taste and attractive flavor. It is expected that passion fruit production and planted area will increase gradually in the years ahead because of high profitability and consumer's demands of healthful ingredients. So we tried to investigate the effect of plant growth regulators and antioxidants on in vitro plant regeneration and callus induction from leaf explants of passion fruit for an establishment of optimal mass propagation system. Young leaf explants of purple passion fruit were cultured in Murashige and Skoog (MS) medium containing different growth regulators and antioxidant additives to induce the shoot organogenesis. After 8 weeks, the highest embryogenic callus formation rate was obtained in MS medium supplemented with $1mg{\cdot}L^{-1}$ 6-benzylaminopurine (BAP) and $2mg{\cdot}L^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), furthermore, the shoot development via organogenesis was also observed. Silver nitrate ($AgNO_3$), which was added into the medium to minimize the adverse effects of leached phenolics, was effective for reduction of medium browning and sudden explant death. In the medium supplemented with $1mg{\cdot}L^{-1}$ BAP and $1mg{\cdot}L^{-1}$ gibberellic acid ($GA_3$), shoots were most vigorously regenerated and elongated. Most shoots rooted successfully in half strength medium with $1mg{\cdot}L^{-1}$ indol-3 acetic acid (IAA), and more than 90% of plantlets survived after 4-month acclimatization period.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.313-317
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• 1997

Leaf segments of the potato (Solanum tuberosum L.) genotypes, ND860-2, Norchip, Russet Norkotah, Goldrush, and Norqueen Russet were transformed with the coat protein gene of potato virus Y (PVY). The white-skinned genotypes, ND860-2 and Norchip, were easily transformed and regenerated into shoots, whereas the three russet-skinned genotypes had low frequencies of regeneration. Transformed shoots were generally recovered in four to six weeks. Antibody to PVY coat protein detected a single band of 30 kD in western blots of transgenic plants. Transformed plants had a normal phenotype in the greenhouse and many showed a delayed buildup of PVY following inoculation. Several transgenic lines had negative ELISA readings 85 days after inoculation. Transgenic lines which did not show detectable levels of PVY antigen will be further tested for resistance to PVY.

• Journal of Ginseng Research
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• v.38 no.3
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• pp.220-225
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• 2014

An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.