• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.88-94
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• 2005

Plant tissue culture studies have been done for the preservation of medicinal plant resources and efficient production of pharmaceutically important secondary metabolites. Micropropagation methods for Cephaelis ipecacuanha have been established and these methods enabled much more efficient propagation of the plants than the conventional methods using seedling or layering. The C. ipecacuanha plants derived from tissue culture grew uniformly in the field and they showed higher alkaloid contents compared to the plants grown from seedlings. Hairy root cultures of C. ipecacuanha and Panax ginseng have been established by infection with Agrobacterium rhizogenes, and the production of important pharmaceuticals by these cultures have been successfully demonstrated. In the case of C. ipecacuanha, the highest alkaloid yields from the hairy roots cultured for 8 weeks were 2.75-fold cephaeline (5.5 mg) and one third emetine (0.7 mg) compared with those from the roots of one-year old plant propagated through shoot-tip culture and cultivated in a greenhouse (2.0 mg cephaeline and 2.0 mg emetine). In the case of P. ginseng, ginsenoside contents in the hairy roots optimally cultured for 4 weeks were much higher than those in the roots of 4-year old field-grown plant. Thus our medicinal plant tissue cultures demonstrate desirable properties. However, they are always exposed to danger of microbial contamination or unexpected trouble of culture facilities. Cryopreservation of plant tissue cultures is a reliable method for long-term preservation. Cryopreservation studies on these cultures are also presented.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.263-270
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• 2006

Pongamia pinnata Pierre is a tree legume, having potential in production of raw material for biodiesel. A protocol for in wk propagation of this plant was standardized using seedling explants. Growth regulators (GR) including gibberellic acid $(GA_3),\;N^6-benzylaminopurine(BA)$, thidiazuron (TDZ), and Adenine sulphate (Ads) were tested for optimum germination of seeds. Removal of seed coat prior to germination, controlled fungal growth partially but enhanced bacterial growth. Antibiotic cefotaxime was ineffective in controlling bacterial contamination. Seedling derived nodal explants and cotyledon nodes with attached cotyledons were excised and cultured for induction of shoots. Optimum sprouting and multiplication of shoot buds were obtained in MS medium supplemented with $8.88{\mu}M$ BA. These buds differentiated and rooted on medium devoid of GR. Optimum growth of Pongamia seedling was obtained in cotton plugged culture vessels. Reculturing of the cotyledon node explants produced more shoots from the same site. This process of removing shoots and reculturing of cotyledon node was followed for eight passages yielding 4 to 8 shoots in each cycle. The shoots (75%) rooted on half strength MS basal medium supplemented with 0.22% charcoal. All plants survived on transfer to soil. This is the first report on in vitro regeneration of Pongamia pinnata. This report demonstrates the possibility of coupling more than one parameter in single experiment to hasten the process of standardization. The process of cycling the nodal explant repeatedly for production of large number of shoots from single meristem may find application in genetic transformation experiments wherein meristems are used for transformation.

• Journal of Ginseng Research
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• v.23 no.3 s.55
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• pp.148-163
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• 1999

The efficacy of three auxins, viz. 2,4-0, NAA and dicamba, were compared for the induction of somatic embryogenesis in American ginseng (Panax quinquefolium L.). Somatic embryos (SEs) formed on ginseng cotyledonary, zygotic embryo and shoot explants after 8 weeks of induction by the auxin stimuli. Significantly more somatic embryos were induced by culture of any of the ginseng explants on media supplemented with $5{\mu}M$ 2,4-0 than any other auxin treatment. Shoots derived from somatic embryos had the greatest regenerative potential and zygotic embryos the least. Explants generated from green (unstratified) seeds gave similar or higher frequency of embryogenesis as the explants derived from stratified seeds. Histological and SEM studies confirmed that the regenerimts were somatic embryos. Somatic embryos germinated and developed into normal plants in $3\~6$ months. About $10\%$ of plantlets from second generation SEs formed flowers within 10 weeks, particularly on media supplemented with $GA_3$ The development of a regeneration system for ginseng through somatic embryogenesis is a necessary first step for mass propagation and genetic improvement of American ginseng.

• Korean Journal of Plant Resources
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• v.35 no.6
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• pp.770-777
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• 2022

The main objective of this study was to identify the most appropriate condition for root formation of in vitro micropropagated pear (Pyrus spp.) plants. In vitro propagation was induced on Murashige and Skoog (MS) medium with 2.0 mg/L of N6-benzyladenine (BA) and 0.2 mg/L of Indole-3-butyric acid (IBA) medium. The short pre-treatment of explants with a high concentration (1 mg/L) of NAA and IBA (R0 medium) in dark for three days, followed by transfer to five different media (R1 to R5) resulted in good rooting responses in the pear 'Oharabani (P. pyrifolia × P. communis)' genotype. For the rooting experiments, the highest rooting percentage (83.3 ± 8.3%), average root length (3.6 ± 1.9 mm), total root number (31 ± 4.0), and average root number per plant (2.6 ± 2.1) were obtained on half strength (1/2) of MS medium supplemented with 30 g/L sucrose without hormones and activated charcoal (AC) (R1 medium). The highest rooting percentage was obtained at 83.3% from explants on R1 and R3 media. The rooting procedure described in this study resulted in good root formation and significantly shorting the root induction time to within 14 days of culture. Further studies are underway to test the suitability of the protocol developed in this study for other pear genotypes.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.382-391
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• 2018

The objective of this study was to determine the most effective medium condition of shoot proliferation and root formation for the efficient in vitro micropropagation of M.26 (Malus pumila Mill). Simple sequence repeat (SSR) markers were used to analyze the genetic diversity of micro-propagated and greenhouse grown M.26. Shoot proliferation was carried out in MS (Murashige and Skoog) containing benzyladenin (BA, $0.5{\sim}5.0mg{\cdot}L^{-1}$) and thidiazuron (TDZ, $0.01{\sim}0.1mg{\cdot}L^{-1}$). The highest number of shoots (10.67 shoots per explant) was induced by adding BA at a concentration $1.0mg{\cdot}L^{-1}$. TDZ treatments caused higher hyperhydricity rate in cultured explants than in BA treatments. There was no significant effect of both BA and auxin on shoot proliferation, and the optimum proliferation medium for M.26 was MS medium containing $1.0mg{\cdot}L^{-1}$ BA. To find a suitable medium composition for shoot rooting, we tested different concentrations indole-3-butyric acid (IBA) and ${\alpha}$-naphthaleneacetic acid ($0.5{\sim}5.0mg{\cdot}L^{-1}$), MS medium (1/4-1), sucrose ($0{\sim}30g{\cdot}L^{-1}$). The shoots showed good rooting on half-strength MS medium containing $1.0mg{\cdot}L^{-1}$ IBA and $15-20g{\cdot}L^{-1}$ sucrose. The rooting rate (100%), number of roots (10.45 ~ 13.60 roots per explant), root length (7.41 ~ 8.33 cm), and shoot length (4.93 ~ 5.38 cm) were good on this medium. Fifteen SSR primers were detected in a total of 30 alleles in 20 micro-propagated plantlets, all SSR profiles from micro-propagated plantlets were monomorphic and similar to greenhouse grown control plantlet M.26 plant. The results indicated that M.26 micro-propagated plantlets were genetically stable.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.484-489
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• 2017

Lilies as cut flowers are one of the most popular ornamental plants in South Korea. It is necessary to develop lily cultivars with high qualities. Therefore, highly efficient propagation systems are needed following release of elite cultivars. In this study, we used taurine treatment to improve the growth conditions including shoot and bulb formation, fresh weight gain, and reduction of rooting and browning. We experimentally evaluated the effect of taurine as a growth stimulator, at concentrations of 0, 2.5, 5, 10, 15 and 20 mg/l. The results showed that 20 mg of taurine enhanced shoot formation by 85% and increased fresh weight 5.5-fold, which was higher than the approximately four-fold increase in the control. In addition, multiple bulb formation rate was increased by 80% and rooting by 82% following exposure to 20 mg/l of taurine. The efficiency of taurine treatment was higher than that of control with 50% multiple bulb formation rate and 60% rooting rate. The browning was 10.6% at 2.5 mg/l of taurine when compared with 0.8% at 20 mg/l. Taurine showed a positive effect on the overall growth of lily plants in terms of increased fresh weight, shoot formation rate, rooting, and formation of multiple bulbs, indicating that taurine can be used as an alternative to amino acids or as an antioxidant such as citrate and vitamin C in plant tissue culture.

• Korean Journal of Medicinal Crop Science
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• v.1 no.2
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• pp.137-157
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• 1993

This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

• FLOWER RESEARCH JOURNAL
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• v.19 no.1
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• pp.8-14
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• 2011

This study was conducted to investigate the effect of age of transplants, propagated by cutting, of two cut rose (Rosa hybrida Hort.) cultivars on their subsequent growth and yield in an effort to develop an efficient cutting propagation method for domestic rose cultivars. Two cultivars used in this study were a standard type 'Pink Aurora' and a spray type 'Yellow King'. Cuttings were prepared as single node cuttings each with a five-leaflet leaf and were stuck in rockwool cubes ($5cm{\times}5cm{\times}5cm$, UR, Korea) at two different dates. Cuttings rooted for either 30 (stuck on Jan. 20, 2009) or 48 days (stuck on Jan. 2, 2009) were transplanted into a rockwool slabs ($10cm{\times}15cm{\times}100cm$, UR, Korea) on the same date, 18 Feb. 2009. Plant growth and cut flower quality were investigated for two successive harvests during the period of Jan. to July in 2009. In both cultivars, 48 days old plants showed some growth of the shoot and root before transplanting. However, in the case of 30 days old plants before transplanting no noticeable growth of the shoot and root was obserable in 'Pink Aurora', while only shoot growth, but not root growth to the bottom of the rooting medium, was observed in 'Yellow King'. This suggested cultivar-specific responses that in this experiment a spray type 'Yellow King' showed greater growth rate during the rooting stage than a standard type 'Pink Aurora'. In the measurement of growth and cut flower yield after transplanting, the 48 days old standard type 'Pink Aurora' produced greater number of cut flowers per plant than 30 days old plants, whereas their mean stem fresh weight was recorded smaller than that of the 30 days old plants. For 'Yellow King', 30 days old plants showed greater stem length, flower width, number of five-leaflet leaves per stem, stem fresh weight, and number of cut flowers per plant than 48 days old plants. Therefore, growth and yield were significantly affected by cultivar and age of the rooted cutting, and additional research is needed on the effect of age of rooted cuttings in more cultivars.

• Journal of Korean Society of Forest Science
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• v.104 no.1
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• pp.76-83
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• 2015

Old growth and protected-trees are worth history and culturally in Korea. This research carried out to investigate habitat characterization and the vegetative propagation methods of Salix chaenomeloides(100-600 years old trees) in Gyeongsangnamdo Province. It has been preserved in good and grow naturally in area surrounding village(4 trees), riverside(1 tree) and field(1 tree). Cutting from old growth and protected trees was conducted using 10 kinds of bedsoils. It was survived above 93% in all bedsoils. Growth of cuttings is a distinct difference according to bedsoils and tree ages. Growth of cutting was high on CBS, followed mixed bedsoils(VPMP) including vermiculite, peatmoss and perlite. Root collar diameter was difference depend on tree ages and bedsoils. All trees are produced 1 to 4 branch. Root growth of cutting was 20 cm on CBS and VPMP, which is the best shoot growing on cutting. Bedsoil with physico-chemical characteristics of high moisturizing ability, high air permeability, high EC value and slightly acid may be suitable for efficiently cutting propagation of old growth and protected S. chaenomeloides. CBS and VPMP bedsoil are favorable for propagating this species by cuttings.

• Korean Journal of Plant Resources
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• v.19 no.4
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• pp.530-536
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• 2006

Hardwood cutting propagation and early growth characteristics were investigated in order to develop the method of cutting propagation and to find out growth characteristics in the low altitude for in situ and ex situ conservation of Empetrum nigrum var. japonicum K. Koch, which is typical arctic alpine plants on Mt. Halla. The growth of roots and shoots was different depending on hormone concentrations or soil conditions. The survival rate, rooting rate, root growth, number of root and shoot growth increased with treatment of 100 mg/l or 500 mg/l NAA. Consequently, optimum condition of hardwood cutting was at treatment with 100 mg/l or 500 mg/l NAA. When plantlets from hardwood cuttings were exposed to the field condition, after 7 months survival rate was 73.3% without shading while $91.1{\sim}94.4%$ at shading conditions. In the green house, however, survival rate of plantlets were $95.6{\sim}97.8%$ without shading. The growth of plantlets was different depending on sites and shading conditions. Particularly, the best growth was obtained when the plantlets were grown in shading conditions. It indicates that relative humidity and light intensity are correlated with the growth in the low altitude area.