• FLOWER RESEARCH JOURNAL
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• v.16 no.4
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• pp.239-246
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• 2008

This study was conducted to establishment of in vitro micropropagation through induction of multiple shoots from axillary bud culture in Calanthe discolor Lindl. Shoots initiation from axillary bud was the most effective on half strength MS medium supplemented with $0.5mg{\cdot}L^{-1}$ NAA and $2.0mg{\cdot}L^{-1}$ TDZ during four weeks of dark culture followed by culture under a 16-h photoperiod. Multiple shoots (12.5 shoots per explant) were proliferated on half strength MS medium containing $4.0mg{\cdot}L^{-1}$ TDZ and $2.0mg{\cdot}L^{-1}$ IBA. On the other hands, the abnormally emerged shoots during the multiple shoot proliferation stage were recovered to normal shoots on half strength MS hormone free medium. Multiple shoots were well elongated and rooted on half strength MS medium with $0.1mg{\cdot}L^{-1}$ NAA. The plantlets were acclimatized up to 100% on TKS substrate after pretreating with $10mg{\cdot}L^{-1}$ NAA for 30 min. and these plantlets showed good growth as well.

• Journal of Bio-Environment Control
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• v.13 no.1
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• pp.51-55
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• 2004

The most commonly used inorganic nutrient compositions such as Murashige & Skoog medium have been optimized for heterotrophic growth. Therefore, they may not be optimal for photomixotrophic and photoautotrophic growth of plantlets. In photomixotrophic micropropagation, emdium sugar level is often lowered, while light and $CO_2$ levels in vessel are raised, and chlorophyllous explants are used to facilitate photosynthetic carbon acquisition. In a factorial experiment effect of addition (+) and omission(_) of organic materials (OM, 0.5 g ${\cdot}$ $m^{-3}$ each of thiamine, nicotinic acid and pyridoxine and 100 ${\cdot}$ $m^{-3}$ myo-inositiol) combined with three sucrose levels (0, 15, and 30 kg ${\cdot}$ $m^{-3}$) was tested on the growth of potato plantlets. Each of nodal cuttings with a leaf was cultured on 0.1${\times}$$10^{-4}m^{-3}$) MS agar ( 8 kg ${\cdot}$ $m^{-3}$) medium (pH 5.80 before autoclave) in glass test tubes (100 mm${\times}$25mm) capped with a sheet of transparent film with a 6 mm diameter gas permeable filter (5.1 air exchanges ${\cdot}$$h^{-1}$). Cultures were maintained in a room for 27 days at $23^{\circ}C$, 50% RH, 350-450${\mu}mol\;{\codt}\;mol^{-1}CO_2$, 16 h ${\cdot}$ $d^{-1}$ photoperiod at 13${\mu}mol\;{\codt}\;m\;{\codt}\;s^{-1}$ PPFD provided by white cool fluorescent lamps. Growth of potato plantlet in the +OM and -OM treatments were similar, while medium pH was 0.2 scale lower in the latter. Dry weight, % dry matter, and stem diameter enhanced, while shoot to root dry weight ratio, leaf area, chlorophyll concentration per gram dry weight, and medium pH decreased with increasing initial sucrose level. Interaction between OM and sucrose levels was observed in shoot length and medium pH. Results indicate that OM can be omitted from the medium without detrimental effect while addition of sucrose was beneficial for the photomixotrophic growth of potato plantlets under raised light and $CO_2$.

• Journal of Plant Biotechnology
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• v.38 no.3
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• pp.221-227
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• 2011

This study was carried out to determine optimal medium conditions for mass propagation by bioreactor culture of ever-bearing strawberry (Fragaria ${\times}$ ananassa D.). Two different type of nitride were that mixed $NH_4NO_3$ and $KNO_3$ or added $KNO_3$ only. And nitride concentrations were at the 4 levels of $1/2{\times}$, $1{\times}$, $2{\times}$ and $3{\times}$ that was included $NH_4NO_3$ and $KNO_3$. Sucrose content ranged at 3 levels of $10g{\cdot}L^{-1}$, $20g{\cdot}L^{-1}$ and $30g{\cdot}L^{-1}$ and medium pH were at the 3 levels of 4.6, 5.6 and 6.6. In bioreactor culture, medium that are included $NH_4NO_3$ and $KNO_3$ together in MS medium was suitable for mass propagation. Medium EC rose rapidly when the nitride concentration was increased. For that reason, plantlet growth was inhibited. Shoots of nitride $1/2{\times}$ concentration was 10.8 ea and $1/2{\times}$ concentration was suitable for shoot propagation. Fresh weight of sucrose $30g{\cdot}L^{-1}$ was 3,101 mg which was heaviest and aerial and ground part were higher than the other concentration. Shoots were increased in proportion to the increasing concentration of sucrose. In the pH condition, from pH 5.6 to 6.8 were appropriate for the optimum growth of aerial and ground part of plant. From the results, in bioreactor culture for mass propagation, MS medium was suitable $1/2{\times}$ concentration that was included $NH_4NO_3$ and $KNO_3$ together, and added $30g{\cdot}L^{-1}$ of sucrose, and then adjusted pH between 5.6 and 6.6.

• Journal of Korean Society of Forest Science
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• v.81 no.3
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• pp.273-279
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• 1992

Protoplasts isolated from leaf mesophyll tissues of Populus koreana ${\times}$ P. nigra var. italica were fused with those of P. euramericana cv. Guardi. Well expended healthy leaves of 5 to 7 week-old-plantlet grown in vitro were used as source materials. Leaves from P. koreana ${\times}$ P. nigra var. italica and P. euramericana cv. Guardi were digested in enzyme solution I (2.0% Cellulase, 1.2% Hemicellulase, 0.4% Macrozyme, 2.0% Driselase, 0.05% Pectolyase ; w/v) and enzyme solution II (1.0% Cellulase, 1.2% Hemicellulase, 0.4% Macrozyme, 2.0% Driselase, 0.05% Pectolyase ; w/v), respectively, The highest frequency of fusion among the protoplasts originated from the two source materials was approximately 21% using 40% PEG or 15% dextran. In addition, fusion frequency was enhanced by incorporating 30mM of $Ca^{2+}$ in eluting solution at pH 10.5. Dividing cells and/or mint-calli were obtained by culturing the fusion products in a liquid 8p-KM medium supplemented with 0.6M sucrose, $0.45{\mu}M$ 2, 4-D, and $0.5{\mu}M$ BA. Shoots were regenerated from the fusion product-derived calli after culture on MS medium containing $5.0{\mu}M$ zeatin. To verify the putative hybrid or cybrid, SDS-PAGE was carried out. From the 24 regenerants, just two plants showed intermediate protein band patterns compared with those of the original source plants.