• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.229-234
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• 2008

An efficient micropropagation was established by using axillary bud explants from two-year-old tree(Echinosphorea koreensis Nakai), which has been known as a rare and endangered species. Among various basal media tested, DKW medium was shown to be the best for axillary shoot elongation. The addition of both BA and TDZ to the medium induced 6 to 10 shoots per explant during eight weeks of culture, without showing any abnormal morphology at the shoot proliferation stage. However, high concentration of TDZ(>0.05 mg/L) appeared to cause hyperhydration on either leaf or shoot at the later developmental stage. Approximately 20% of shoots produced roots by the addition of 1.0 mg/L NAA but not by IBA($0.2{\sim}1.0$ mg/L). Ex vitro micro-cuttings were better source for root induction; up to 58.6% of the micro-cuttings rooted when 100 mg/L IBA was applied to the soil(vermiculite). More than 90% of plantlets with roots were successfully acclimatized and grew normally in the field. Therefore, we suggest that this endangered tree species can be effectively micropropagated by axillary bud culture system developed in this study.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.257-262
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• 2005

Agrobacterium tumefaciens-mediated cotyledonary-node explants transformation was used to produce transgenic melon. Cotyledonary-node explants of melon (Cucumis melo L. cv. Super VIP) were co-cultivated with Agrobacterium strains (LBA4404, GV3101, EHA101) containing the binary vector (pPTN289) carrying with CaMV 35S promoter-gus gene as reporter gene and NOS promoter-bar gene conferring resistance to glufosinate (herbicide Basta) as selective agent, and the binary vector (pPTN290) carrying with Ubiquitin promoter-GUS gene and NOS promoter-nptll gene conferring resistance to paromomycin as selective agent, respectively. The maximum transformation efficiency (0.12%) was only obtained from the cotyledonary-node explants co-cultivated with EHA101 strain (pPTN289) on selection medium with 5 mg/L glufosinate and not produced a transgenic melon from the cotyledon or cotyledonary-node co-cultivated with other strains. Finally, five plants transformed showed the resistance in glufosinate antibiotic and the GUS positive response in leaf ($T_0$), flower ($T_0$), seeds ($T_1$) and plantlet ($T_1$). Southern blot analysis revealed that the gus gene integrated into each genome of transgenic melon.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.301-306
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• 2005

Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

• Journal of Plant Biotechnology
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• v.31 no.1
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• pp.89-94
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• 2004

Callus of Rhodiola sachalinensis A. Bor were induced from leaf explant on l/2MS solid supplemented with combination of auxin (2.4-D, NAA: 0.1∼2mg/L) and cytokinin (BA: 0.1∼0.2mg/L). The effects of various medium, culture conditions and phytohormones on the growth of callus were investigated. MS, WPM, B$_{5}$ medium and diluted or concentrated media (1/2X, 2X, 3X) were used to investigate the growth of callus on each media. Among these, the highest growth was observed when cultured in in 2B$_{5}$ medium containing 0.5mg/L NAA and 1mg/L BA, 3％ sucrose and 49.6 mM KNO$_3$ as nitrogen source, and 2.16mM NaH$_2$PO$_4$ as phoshate source at $25^{\circ}C$ in the dark. The calli cultured with 5％ sucrose produced high salidroside content (0.41％ on the basis of dry wt) than normal root (0.17％).

• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.259-264
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• 2002

Induction of embryogenic callus from Allium wakegi Araki explants was promoted on medium containing 2,4-D, and production of abnormal embryos from the embryogenic callus increased as 2,4-D concentration was raised. Shoot tip was found to be the best explant source for embryogenic callus formation followed in the order by bulb scale and leaf section. Medium containing 0.09 M sucrose was effective for embryogenic callus production. The regenerants from embryogenic callus on medium containing 2,4-D and BA at different concentrations was consisted of diploids, tetraploids and a few mixaploids of 2n＋4n, and their chromosomal aberration rate ranged from 8.0 to 33.3%. Frequency of chromosomal aberrants was the highest (18.7%) in the regenerants obtained from bulb scale-derived embryogenic callus among others. Plant regeneration rate was high (33.5%) from the shoot tip-derived embryogenic callus and the frequency of chromosomal aberrants was very low (7.0%). The plantlets regenerated on medium containing 0.26 M sucrose resulted in high chromosomal aberrants. The regenerants on medium containing sucrose at 0.09∼0.20 M produced chromosomal aberrants at around 15.2∼16.6%.

• Korean Journal of Plant Resources
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• v.20 no.4
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• pp.367-374
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• 2007

This study was carried out to establish the micropropagation system of Bupleurum latissimum Nakai that is a Korean native endangered species. Callus were induced from the leaf, petiole and floral bud and the percentage of callus formation was highest in the floral bud on the MS medium containing 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D. Especially, callus induced from floral bud was formed 77.8% and the percentage of shoot formation was 42.6% on the MS medium containing 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D plus 1.0 mg ${\cdot}$ $L^{-1}$ TDZ. For simultaneously callus formation and shoot regeneration, 1/2 MS medium was more effective than MS medium. The percentage callus formation, shoot regeneration and rooting were 46.3%, 13.0%, 13.0% in 1/2 MS medium, respectively. Soot regeneration from callus was good in 1/2 MS medium supplemented with 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D plus 1.0 mg ${\cdot}$ $L^{-1}$ BA where percentage of shoot regeneration was 74.1 %, and the number of shoot per explant was 2.4. The percentage of rooting was lowest (57.8%) in control while it was highest (97.8%) in 1.5 mg ${\cdot}$ $L^{-1}$ NAA. In acclimatization of regenerated plantlets, the percentage of survived plantlets was highest (86.1%), and plant height, root length and fresh weight were good in the soil for horticulture.

• Journal of Plant Biotechnology
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• v.39 no.1
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• pp.69-74
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• 2012

Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11a
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• pp.29-30
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• 2002

The goal of this research was to develop the effectiveness of in vitro culture method for A. formosanus and study the environment in vitro conditions affecting on growth. The first series of experiments were examined to investigate the response of three different basal media, MS (Murashige and Skoog, 1962), Knudson (KC; Knudson, 1946) and modified hyponex on growth and multiplication during in vitro culture. Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with BA (1 mg1$\^$-1/) or TDZ (1-2 mg1$\^$-1/). Addition of activated charcoal (1%) to the TDZ containing medium promoted rapid shoot tip proliferation (11.1 shoots per explant) but the same medium had an opposite effect resulting in poor proliferation in the nodal explants. However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 media supplemented with 2% sucrose and 0.5% activated charcoal. Using bioreactor culture for scaling up was also shown the best way for multiple shoot induction and growth of this plant. The second series of experiments was studied to investigate the effect of physical environment factors on growth of in vitro plantlets. The Anoectochilus formosanus plantlets were cultured under different air exchange rate (0.1, 0.9, 1.2h$\^$-1/), without sucrose or supplement 20g.1$\^$-1/ (photoautotrophic or photomixotrophic, respectively), and different photosynthesis photon flux (40, 80, 120 ,${\mu}$mol.m$^2$.s$\^$-1/- PPF). Under non-enrichment CO$_2$ treatment, slow growth was observed in photoautotrophical condition as compared with photomixotrophical condition on shoot height, fresh weigh and dry weight parameters; High air exchange (1.2.h-l) was found to be inadequate for plant growth in photomixotrophical condition. On the contrary, under CO$_2$, enrichment treatment, the plant growth parameters were sharply (visibly) improved on photoautotrophic treatments, especially on the treatment with air exchange rate of 0.9.h-1. The growth of plant in photoautotrophic condition was not inferior compared with photomixotrophic, and the best growth of plantlet was observed in treatment with low air exchange rate (0.9.h-1). Raising the PPF level from 80 to 120${\mu}$mol.m$\^$-2/.s$\^$-1/ decreased the plant height, particularly at 120${\mu}$mol.m$\^$-2/.s$\^$-1/ in photoautotrophic condition, fresh weight and dry weight declined noticeably. At the PPF of 120${\mu}$mol.m$\^$-2/,s$\^$-1/, chlorophyll contents lowed compared to those grown under low PPF but time courses of net photosynthesis rate was decreased noticeably. Light quality mainly affected morphological variables, changes of light quality also positively affected biomass production via changes in leaf area, stem elongation, chlorophyll content. Plant biomass was reduced when A. formosanus were grown under red LEDs in the absence of blue wavelengths compare to plants grown under supplemental blue light or under fluorescent light. Stem elongation was observed under red and blue light in the present experiment. Smaller leaf area has found under blue light than with other lighting treatments. Chlorophyll degradation was more pronounced in red and blue light compared with white light or red plus blue light which consequent affected the photosynthetic capacity of the plant. The third series of experiment were studied to investigate the effect of physical environment factors on growth of ex vitro plants including photosynthesis photon flux (PPF), light quality, growing substrates, electrical conductivity (EC) and humidity conditions. In the present experiments, response of plant on PPF and light quality was similar in vitro plants under photosynthesis photon flux 40${\mu}$mol.m,$\^$-2/.s$\^$-1/ and white light or blue plus red lights were the best growth. Substrates testing results were indicated cocopeat or peat moss were good substrates for A. formosanus growth under the greenhouse conditions. In case of A. formosanus plants, EC is generally maintained in the range 0.7 to 1.5 dS.m-1 was shown best results in growth of this plant. Keeping high humidity over 70% under low radiation enhanced growth rate and mass production.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.1
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• pp.110-114
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• 1981

These experiments were carried out to define the effects of NAA, 2, 4-D and Benzyladenine on the callus induction and the organ differentiation from the explants and to find out the vegetative propagation method of Korean ginseng. The results obtained are summarized as follows; 1. NAA was significantly effective in forming roots from the ginseng stem segment and the number of roots was increased by increasing NAA concentration in the medium. The roots were formed from both distal and proximal ends of the ginseng stem segments grown on the medium containing more than 2mg/L of NAA. 2. The amount of callus growth increased proportionatly with NAA concentration in the range of 4.0mg per liter in the medium. The callus was easly induced from stem segment than leaf segment and 2, 4-D was more effective in callus induction and growth than NAA. 3. The benzyladenine showed the significant inhibition effect in forming roots from ginseng explant. The callus was not induced with BA alone, but in BA and 2, 4-D or BA and NAA added medium, the callus was easily induced and its growth was also accelerated. The interaction effects between 2, 4-D and BA on the callus induction and growth were significantly higher than those between NAA and BA. 4. As the ginseng embryos were cultured on the M.S. medium supplemented with 2mg per liter NAA, number of shoots was significantly increased and the percentage of embryo which had shown more than 4 shoots later was 22.2%. On the medium containing 8mg per liter NAA, the ginseng embryo showed the normal growth of shoots and leaves, but increased roots and callus induction on the basal part of shoots. 5. When the shoots with 3 leaflets were cut in 1.5cm long and grown on the Blayde's medium containing NAA 1.0mg per liter, roots were formed at the proximal end of shoot, and a new ginseng seedling was successfully obtained.