• Journal of The Korean Society of Grassland and Forage Science
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• v.28 no.3
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• pp.171-176
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• 2008

Efficient plant regeneration system of birdsfoot trefoil (Lotus corniculatus L.) was development. The factors affecting the somatic embryo formation, its proliferation and regeneration capacity of leaf and stem explants of Empire cultivar was investigated. The highest number of somatic embryos was obtained on B5 medium supplemented with 1 mg/L NAA and 1 mg/L BA. Depending on different explants, highest frequency of embryogenic callus and regeneration were observed in Empire with leaf explants. The response from stem explants was slower and callus induction was less than that from leaf explants. Regenerated shoots formed complete plantlets in on 1/2 MS medium supplemented with 1 mg/L IBA. Regenerated plants were morphologically uniform with normal shape and growth pattern.

• Journal of Korean Society of Forest Science
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• v.77 no.2
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• pp.208-215
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• 1988

The leaf mesophyll protoplasts of Populus alba ${\times}$ glandulosa were isolated from leaf of plantlet in vitro and cultured for plant regeneration. The MS medium (minus $NH_4NO_3$) with 0.5 mg/l BAP and 2.0 mg/l 2, 4-D showed the moderate frequency of dividing protoplasts cultured by the liquid plating method during the first week of culture. The percentage of colony formation was revealed the highest frequency by the gauze contained semi-solid agar plating method after 5 weeks cultured. Ridding out the gauze, the micro-callus was formed on the same semi-solid medium in 8 weeks after protoplasts culture. For proliferation of callus, mini-callus was transferred on the MS solid medium with 0.5 mg/l 2, 4-D and 0.1 mg/l BAP 12 weeks after culture. Shoot regeneration occurred when the calli derived from protoplasts were cultured on MS medium with 1.0 mg/l zeatin and such shoots could be readily rooted on the one half strengthen MS medium with non-phytohormone. Rooting shoots were planted in green-house 22 weeks after protoplast culture.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.206-209
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• 2001

Recombinant human GM -CSF was expressed and secreted from transgenic rice cell suspension cultures in its biologically active form. This was accomplished by transforming rice callus tissues with an expression vector, pMYN44. containing the hGM -CSF cDNA. Regulated expression and secretion of hGM -CSF from this vector achieved using the promoter, signal peptide, and terminator from a rice alfa-amylase gene Amy3D. The Amy3D gene is expressed in response to sugar deprivation. The recombinant hGM -CSF was expressed from the transgenic rice cell culture on the sugar-free medium as a yield of about 110 mg/L in the culture filtrate, which was determined by ELISA. Biological activity of hGM-CSF was confirmed by measuring the proliferation of the hGM -CSF dependent TF -1 cells.(This work was supported by a grant from the NRL program of the Korean Ministry of Science and Technology. Shin, Y.- J.. Lee. J.-H and Kwon, T.-H. have been supported by BK21 program from the Korean Ministry of Education)

• Plant Biotechnology Reports
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• v.2 no.3
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• pp.179-189
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• 2008

We established an efficient plant regeneration system for Catharanthus roseus L. (G.) Don through somatic embryogenesis. Embryogenic callus was induced from hypocotyl of seed germinated in vitro. Somatic embryogenesis in Catharanthus has been categorized into three distinct stages: (1) initiation and proliferation of embryo; (2) maturation, and; (3) germination or plantlet conversion. Beside plant growth regulators, various stages of embryogenesis were screened for their response to a wide variety of factors (pH, gelrite, light, sugar alcohols, polyethyleneglycol and amino acids), which affect embryogenesis. All of the tested factors had a small to marked influence on embryogeny and eventual conversion to plantlets. The plantlets were acclimatized successfully in a greenhouse. To our knowledge, this is the first report describing a detailed study of various cultural factors which regulate embryogenesis in C. roseus. The results discussed in this paper may be used in mass propagation to produce medicinal raw material, and the embryo precursor cells could be used in genetic modification programmes that aim to improve the alkaloid yield as well.

• Journal of Korean Society of Forest Science
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• v.97 no.4
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• pp.452-457
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• 2008

In order to develop an efficient micropropagation technique effect of plant growth regulators (PGRs) affecting on shoot proliferation from shoot apex in Pyrus ussuriensis was tested. Generally, there was no conspicuous effect on shoot induction by the treatment of PGRs and one or two shoots/explant were induced when cultured on MS medium supplemented with BA and/or BA plus NAA. Both apical shoot necrosis and hyperhydric shoots were observed frequently in multiplied shoots, and callus was formed at the basal part of shoots. About 20% spontaneous rooting was achieved in growing shoots, however the proliferated shoots exhibited poor rooting rate in gelrite supported media. When we tried to ex vitro rooting of the shoot cutting, the shoot cuttings rooted up to 50% with 100 mg/L IBA application. The rooted plantlets grew normally after acclimatization in the greenhouse.

• 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.

• Korean Journal of Plant Resources
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• v.32 no.1
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• pp.53-62
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• 2019

The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.

• Journal of Bio-Environment Control
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• v.31 no.2
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• pp.133-141
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• 2022

Bulbophyllum auricomum Lindl. is a rare orchid and has flowers with an attractive fragrance. The present study investigated the tissue culture method for micropropagation. Capsules derived from artificial self-pollination were obtained for the best seed germination in MS basal medium. Plant growth regulators (1.0 mg·L^-1 of BAP and 2.0 mg·L^-1 of NAA) were affected by callus induction from subcultured pseudobulb explants. For the callus subculture, different natural plant extracts were tested in 11 treatment media. Among them, MS medium with 150 mL·L^-1 of coconut water was generally effective in fresh weight (1.75 ± 0.08) and (3.01 ± 0.20) of callus proliferation and PLBs induction at 1 and 2 months, respectively, followed by an MS combination of 30 g·L^-1 of banana and 20 g·L^-1 of potato extract. The results of a comparative study of different MS mediums containing plant growth regulators with a natural extract combination and MS medium supplemented with natural extract only showed that MS medium supplemented with a combination of natural extracts (150 mL·L^-1 of coconut water) and plant growth regulators (2.0 mg·L^-1 of BAP and 1.0 mg·L^-1 of NAA) obtained the highest shoot regeneration (3.37 ± 0.17) and (6.41 ± 0.68) after 1 month and 2 months of culturing, respectively.

• Korean Journal of Medicinal Crop Science
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• v.7 no.2
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• pp.94-100
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• 1999

To establish the mass propagation system of Lavandula spica cv. Marino, shoot tip, node, internode and leaf segment cultures were carried out. RAPD was applied to detect the somaclonal variation. Callus induction was very high in the medium supplemented with 1 mg/l 2.4-D, 2 mg/l NAA. especially and combined with 0.05 mg/l BAP from leaves. Shoot formation was high with $2{\sim}4\;mg/l$ BAP or 4 mg/l BAP + 0.2 mg/l NAA from shoot tip. Shoot proliferation was 9.1 times in the $B_{5}$ medium with 0.5 mg/l BAP and 0.01 mg/l NAA. Root formation was improved in NAA, which was the concentration of 0.1 to 1 mg/l and 1 mg/l IAA. Nursery survival rate was enhanced over 90% and growth was looked good in the acclimation soil consisting of peatmoss : vermiculite : perlite (1:1:1, v:v:v). Randomly amplified polymorphic DNA banding patterns based on polymerase chain reaction (PCR) were used to assess the genetic variation in plants regenerated from in vitro culture.

• Journal of Plant Biotechnology
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• v.36 no.1
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• pp.7-12
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• 2009

This study was conducted to establish the optimal suspension culture system for both the propagation of embryogenic cells (ECs) and the induction of somatic embryos (SEs) of Kalopanax septemlobus. The proliferation rate of ECs was reduced as the inoculum density was increased; the highest rate was obtained when 0.1 g/100 ml of cells was initially inoculated. According to the analysis of cell growth pattern and cell growth cycle (G1, Sand G2/M), the cell growth started in 5 days culture initiation, grew rapidly until 15 days and then decreased gradually. Distinctive changes of the cell growth cycle by the culture periods was also observed; the growth cycle was doubled from initial 5.6% to 11.7% of S stage in 5 days culture and then reached in stable stages again. Therefore, the results indicated that a 15-day-cycle was the optimal culture period for the propagation of the ECs through the suspension culture. Furthermore, the cell inoculum density was also important for the induction of SE; more than 65% of SEs at the torpedo stage was induced by using the low level of cell inoculum (0.5 g/L), while the higher inoculum densities were rapidly reduced the proportion of SEs at that stage. Although the higher inoculum density delayed the development of SE, it did not affect the proportion of SEs at the globular and heart stage. In conclusion, this study showed that the suspension culture of the Kalopanax septemlobus ECs through the control of inoculum density was an efficient way for both the propagation of ECs and the induction of SEs, suggesting that the development of this system might help to reduce the culture period for the somatic embryo production.