• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.375-380
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• 2003

This study was conducted to establish a practical masspropagation system of Phalaenopsis clones from basal shoot segments. The frequency of PLB (protocorm like body) induction was compared with various explants. Basal shoot segments showed the most successful result of 45％, while root tips, stalk node segments, stalk leaves and mature leaves represented low frequency (below 5％). The PLB induction ratio in the culture of basal shoot segments was examined with 11 different Phalaenopsis varieties, and the majority of varieties, including pink flower lines, showed an about 30％ rate of PLB formation. Especially, when whole basal shoot parts without cutting were inoculated onto PLB induction medium, giant PLB was induced from explant. This giant PLB was green color and big in size compared with normal PLB. When dissected giant PLB segments inoculated onto PLB multiplication medium, only normal size of PLBs were induced from them. PLBs induced by basal shoot culture were transferred onto proliferation medium and then shooting medium, from which normal plants were formed. Therefore, this culture method is considered as effective and practical protocol for Phalaenopsis mericlone production. In addition, it is suggested that clones of an infinite number can be produced consecutively by this culture system through repeated cycles of PLB induction and proliferation using the basal shoot segment of flask plant.

• FLOWER RESEARCH JOURNAL
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• v.17 no.1
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• pp.9-14
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• 2009

This study was conducted to find out the commercial in vitro propagation methods through protocom-like bodies (PLB) induced from shoot tip culture of Miltonia spp. Among several culture media for induction PLB from shoot tip in Miltonia spp., MS basal medium was better than Hyponex, Vacin & Went basal medium and other media supplemented with natural additives. PLB's proliferation and differentiation in Hyponex medium including $20g{\cdot}L^{-1}$ banana + $20g{\cdot}L^{-1}$ sucrose(pH 5.2) was better than in MS medium. It was tendency that solid media showed higher PLB fresh weight than liquid medium or other cotton bridge culture. The dark culture for 1~2 weeks and adding $10{\sim}20g{\cdot}L^{-1}$ sucrose on hyponex basal medium was the most effective to increase the PLBs growth and shoot number.

• Journal of Forest and Environmental Science
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• v.30 no.1
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• pp.126-132
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• 2014

This experiment was conducted to establish the in vitro propagation of Dendrobium moniliforme through protocorm like body (PLB) induction from the culture of leaf, stem and root explants. Frequency of PLB formation from root explants was higher than those of leaf and stem explants. PLB's proliferation in MS medium including 1.0 mg/L NAA was better than 1.0 mg/L IBA. Frequency of PLB's proliferation on medium with various concentrations of BA, Kinetin, Tidiazuron (0. 1.0, 3.0 mg/L) and NAA (0, 1.0 mg/L) was tested. The maximun proliferation of PLB's was obtained on MS medium supplemented with 3.0 mg/L BA and 1.0 mg/L NAA after 6 weeks of culture. Addition of 200 mg/L activated charcoal (AC) and 30 g/L sucrose was effective on PLB proliferation for D. moniliforme.

• KSBB Journal
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• v.5 no.1
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• pp.43-47
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• 1990

It was determined optimal Culture conditions and suitable growth regulators for seed germination, growth of callus, and protoplasts derived from cultured and mesophyll cells in Gremastra appendiculata. Induction of fusion between protoplasts of cultured and mesophyll cells was examined. The best conditions of seed germination and growth of callus were achieved on Hyponex medium contained plant growth regulators(2mg/l 2, 4-D, lmg/l Kinetin). Viability and regeneration of cell wall in protoplasts was determined with fluorescence microscope. Also, fused protoplasts were achieved by using PEG solution between protoplasts of cultured and mesophyll cells.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.900-910
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• 2015

This study aimed to investigate the efficiency of colchicine and oryzalin in inducing polyploidy in two Cymbidium hybrids [Showgirl 'Silky' and Mystery Island 'Silk Road' (Silk Road-4)]. Colchicine was used at concentrations ranging from 50 to $500mg{\cdot}L^{-1}$, with treatments lasting 1 to 3 weeks. Oryzalin was used at concentrations ranging from 3 to $20mg{\cdot}L^{-1}$, with treatments lasting 3 to 6 days or 1 to 3 weeks. The survival rate of PLBs was better in colchicine than in oryzalin solutions. The ploidy levels were screened using flow cytometry. In C. Showgirl 'Silky', the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (60%) and the 2-week treatment in $5mg{\cdot}L^{-1}$ oryzalin (46.7%). In C. Mystery Island 'Silk Road' (Silk Road-4), the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (16.7%) and the 3-day treatment in $10mg{\cdot}L^{-1}$ oryzalin (6.7%). Colchicine was more efficient than oryzalin in terms of polyploidy induction. Furthermore, pre-treatment, which entailed poking 10 times with forceps, improved the efficiency of chromosome doubling.

• Horticultural Science & Technology
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• v.29 no.3
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• pp.260-266
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• 2011

An efficient protocol was established for high frequency somatic embryogenesis through a callus culture of Coelogyne cristata. The best frequency of callusing was obtained from a PLB segment (3-5 mm) cultured on MS medium supplemented with coconut water (CW) and a combination of both 3 $mg{\cdot}L^{-1}$ of 2,4-D and BA. When the calli were sub-cultured on the MS medium without any PGRs, the average number of somatic embryos were higher than those with PGRs treatment. NAA is the most critical factor among PGRs, which dramatically hindered for the formation of a somatic embryo. The efficacy of the addition of coconut powder (CP) for somatic embryogenesis was almost the same in all treatments. However, the number of somatic embryos formed distinctly depended on age of the callus. The somatic embryos converted into healthy plants with well-developed shoots on the same medium. Plantlets showed the best responses of root and shoot growth when transferred to $\frac{1}{2}$ MS medium containing 1.5 $g{\cdot}L^{-1}$ of activated charcoal. All plants with above 3.0-cm-high were successfully acclimatized in the greenhouse.

• 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.