• The Journal of Natural Sciences
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• v.4
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• pp.143-159
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• 1991

In order to investigate the micropropagation of Pelargonium, 2 cultivars of P. peltatum 'Pouletta' and P. zonale 'Pinto Red' were cultured in vitro on the MS basal medium supplemented with various concentrations of growth regulators. It attempted to study the induction of callus and the differentiation of organs from leaf disc, petiole segments, stem segments. hypocotyle segments and flower stalk segments. The results are summarized as follows; A. As for the initiation of callus, stem explant was proved to be the most suitable one among various explants of P. zonale 'Pinto Red'. The medium was supplemented with 1.0mg/1 BAP and 1.0mg/1 NAA. As NAA concentration increased, callus formation was enhanced, but higher concentration of NAA inhibited callus fromation. Leaf and hypocotyle explants showed less callus formation than stem and petiole explants. B. In P. zonale 'Pinto Red' petiole culture, the condition of cullus culture such as hormone concentration resulted in affecting shoots differentiation. The best result of shoots formation from the callus reculture were obtained from the combination of 0.5-1.0mg/1 BAP and 0.1-1.0mg/1 NAA when the callus was cultured in 1.0mg/1 BAP and 0.05mg/1 NAA. When the callus was cultured in medium without BAP, the shoot was not differentiated in subculture regardless to BAP and NAA concentration. and only callus was formed. C. Poly-phenol substance was observed in MS medium supplemented without PVP, in which callus was not formed from the leaf of P. peltatum 'Rouletta'. Polyphenol substance was not observed in MS medium supplemented with PVP, in which callus formation was increased. D. The callus formation of P. peltatum 'Rouletta' showed the stem explant being best result. The best result particularly in the stem explant among others. The optimal hormonal concentration was 0.1mg/1 NAA and 5.0mg/1 BAP. The shoot formation was observed at 0.05mg/1 NAA and 1.0mg/1 BAP, 0.1mg/1 NAA and 5.0mg/1 BAP. The shoot was malformed and the tissue recultured turned necrotic.

• 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.

• Journal of Plant Biotechnology
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• v.46 no.2
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• pp.127-135
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• 2019

The purpose of this study was to investigate suitable parts for callus induction and optimal concentrations of growth regulators, contained in the medium affecting shoot and rooting for in vitro mass production of Haworthia truncata. Leaves and flower bud showed 100% callus formation rate at NAA $1{\sim}2mgL^{-1}$ treatment, and NAA $1mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment. The flower stalk showed 75% callus formation rate, at NAA $2mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment in H. truncata. While the rate of callus formation was high in leaves and flower bud, leaves were the most efficient in obtaining most culture parts. Shoot induction rate from callus was highest, at NAA $0.1mgL^{-1}$ treatment in H. truncata. Additionally, the number of shoots formation was 66.3 shoots high, in NAA $1mgL^{-1}$ + BA $0.1mgL^{-1}$ treatment in H. truncata. In the case of acclimatization of regenerated plant, growth characteristics did not show significant difference (95%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate, considering plant height and appearance preference of H. truncata. It was established that optimization of culture condition, was responsible for mass propagation in vitro cultures of H. truncata.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.37-37
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• 2021

Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L^-1, and IBA 1mg·L^-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

• Journal of Plant Biotechnology
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• v.45 no.2
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• pp.110-116
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• 2018

This study was conducted to develop a simple, rapid, and reliable method for in vitro propagation of disease-free and true-to-type clones from sucker explants of thornless blackberry (Rubus fruticosus L. ${\times}$ R. parvifolius L.). To induce multiple shoots, the sucker explants were sterilized in 1% NaOCl solution, and then were aseptically cultured on the full and 1/2 MS solid medium supplemented with BAP (0.1, 0.5, 1.0, 2.0 mg/L). After six weeks of culture, the highest frequency (85.4%) of shoot formation from sucker explants was obtained on the full-strength MS medium with 1.0 mg/L BAP. Node explants obtained from multiple shoots were cultured on the various media of full- or half-strength of AD, B5, MS, SH, QL, WPM media, respectively. After 30 days of culture, plant growth was good on the half-AD, half-QL medium. After 90 days of culture, plant growth was good on the full MS and full SH medium. The survival rate of the plantlets after transfer to plastic pots containing soil mixture (sand: soil: vermiculite was 1:1:1, vol.) in the greenhouse was 98%. The results indicate that a multiple-shoot procedure can be applied for an efficient mass propagation of Rubus fruticosus L. ${\times}$ R. parvifolius L.

• Korean Journal of Plant Resources
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• v.18 no.3
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• pp.382-389
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• 2005

This study was conducted to establish the system of effective in vitro propagation by various explant sources culture of Bippeastrum hybridum Hort, 'Dazzler'. We tested the effects of optimal explant source, plant growth regulators on bulblet formation and plant regeneration. Callus was readily produced on the different tissues excised from floral buds whereas, bulbs and shoots were formed only on pedicel explants as compared with anthers, styles and ovaries. Pedicel is the best optimal explant for in vitro propagation. Two distinct pathways, organogenesis through callus and direct bulblet formation, could be recognized in pedicel culture. Up to the $80-100\%$ of bulblet formation and shoot organogenesis from the pedicel in fifteen days before anthesis were effectively induced by MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA. Plantlet regeneration was successfully achieved from pedicel-derived callus, via shoot bud induction or direct bulblet formation. The bulblets with blooming flower were produced within 2 years.

• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.222-229
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• 2022

Apples are the most grown fruit crops in the fruit industry of Korea. However, virus or viroid infection such as apple mosaic virus (ApMV), apple stem grooving capillovirus (ASGV), apple stem pitting virus (ASPV), apple chlorotic leaf spot virus (ACLSV), apple scar skin viroid (ASSVd) causes fruit yield reduction and poor fruit quality. Therefore, in this study, we examined to established an efficient virus-free system to eliminate the most infected ASGV virus in domestic apple orchard. We investigated that the shoot growth rate and the virus removal rate in ASGV infected potted apples that were treated with heat treatment in a growth chamber (constant temperature/humidity device) maintained at 36℃, 38℃ and 40℃ for 4 weeks. Here we found that the shoot growth rate was the highest in the heat treatment group (36℃) and the virus was removed in the middle and top of the shoot but not in the bottom. The virus was did not removed in the 38℃ and 40℃ heat treatment group in all section of shoots, and the heat treatment group (40℃) died after 4 weeks of heat treatment without growth of shoots. We performed in vivo shoot-tip grafting using the shoot-tip of potted apple heat-treated at 36 ℃, and we also investigated the viability and virus removal rate, which showed 94% viability and 20% virus removal rate. Collectively, our results suggest that it would be possible to produce the virus-free apple plants through heat treatment and shoot-tip grafting.

• Horticultural Science & Technology
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• v.19 no.1
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• pp.17-21
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• 2001

To develop a selection system for regenerating plants from transformed tissues, effects of four antibiotics (kanamycin, hygromycin, carbenicillin, cefotaxime) and herbicide (phosphinotricin) on shoot regeneration from cotyledon and hypocotyl explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis) were studied. For cotyledon, shoot induction was not significantly affected by kanamycin at $1mg{\cdot}L^{-1}$, but the number of shoots formed was significantly reduced at $2mg{\cdot}L^{-1}$, and no shoots were regenerated from any explants at $6mg{\cdot}L^{-1}$ or higher. Hypocotyl explants showed similar result as cotyledon. Kanamycin at $7mg{\cdot}L^{-1}$ may be adequate for selecting Chinese cabbage transformants. Hygromycin at $4mg{\cdot}L^{-1}$ or higher completely inhibited the growth and shoot regeneration of Chinese cabbage explants. Therefore, resistance gene to hygromycin may also be used as a selective marker for Chinese cabbage transformation. Carbenicillin and cefotaxime, the cephalosporin type of antibiotics, had little effect on shoot regeneration of Chinese cabbage explants. Since carbenicillin and cefotaxime have low toxicity to Chinese cabbage, they are suitable for use in tissue culture to eliminate Agrobacterium in transformation experiments after co-cultivation. Shoot regeneration from cotyledon and hypocotyl explants was significantly reduced in presence of $1mg{\cdot}L^{-1}$ phosphinotricin (PPT) and completely inhibited by $2mg{\cdot}L^{-1}$ or higher. PPT, same as antibiotics, may also be used to select transformed cells. Since Chinese cabbage is known to be recalcitrant to in vitro shoot regeneration compared to other Brassica species, even though lower levels of selectable markers result in more transformants but simultaneously allow more untransformed escapes to develop, lower levels of antibiotics and herbicides could be successfully used as a selectable marker to reduce selection pressure.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.163-169
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• 1999

This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.129-135
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• 1997

Morphogenetic responses were investigated by culturing embryo and leaf explants of Korean wild type tea plant, Camellia sinensis (L.) O. Kuntze. Induction of direct somatic embryogenesis as well as adventitious and/or axillary shoots was obtained from mature zygotic embryo cultures on Murashige and Skoog (MS) basal medium having 5 to $20\mu\textrm{M}$cytokinin a lone. Morphogenetic response was decreased dramatically by the addition of auxins tested. One hundred percent of induced and isolated shoots formed roots after four weeks of culture on half-strength MS or quarter-strength Schenk and Hildebrandt (SH) media supplemented with $10\mu\textrm{M}$indole-3-butyric acid (IBA). Immature zygotic embryos were shown to be a suitable explant for embryogenic callus formation in the presence of 2, 4-dichlorophenoxyacetic acid(2, 4-D) in basal medium. Mature zygotic embryo originated leaves were used to test their ability for mophogenesis by incorporating plant growth regulators such as IBA, naphthyl-1-acetic acid (NAA), and 6-benzylaminopurine (BAP). Apparently, the morphogenetic responses of the cultured explant sources on the types and/or levels of plant growth regulators tested were observed visually.